The present invention relates to recombinant DNA comprising DNA coding for smooth-muscle-type myosin heavy chain SM1 isoform protein inserted into vector DNA, a microorganism carrying the recombinant DNA, and an agent for treatment of arteriosclerosis comprising the recombinant DNA. The recombinant DNA of the present invention can be used effectively as an agent for gene therapy of restenosis after PTCA treatment.

Description

22 parts

›TECHNICAL FIELD

The present invention relates to recombinant DNA comprising DNA coding for myosin heavy chain SM1 isoform protein inserted into vector DNA, a microorganism carrying the recombinant DNA and an agent for treatment of arteriosclerosis comprising the recombinant DNA which are used in gene therapy.

›BACKGROUND ART

Smooth muscle-type myosin heavy chain SM1 isoform protein is responsible for contraction and relaxation of smooth muscles, and is one of the myosin heavy chain isoform proteins expressed specifically in smooth muscle cells. As the DNA coding for the protein, the nucleotide sequence of cDNA coding for rabbit SM1 isoform is known (P. Babij et al.: Proc. Natl. Acad. Sci. USA, 88, 10676 (1991)), but there is not known any homology among nucleotide sequences for such DNAs. Further, there is not known any recombinant DNA comprising DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein inserted into vector DNA which recombinant DNA can be injected into animal cells.

It has been reported that when cDNA coding for a protein called calponin which is a protein existing in smooth muscle cells is introduced and expressed in a vascular smooth muscle cell line derived from rat pulmonary arteries, the time required for doubling the cells is prolonged by about 4 hours (Takahashi et al.: Circulation, 88, I-174 (1993)) and that when cDNA for human calponin is injected into a rabbit topically at a site where the carotid artery has been abraded with a balloon, thickening of the intima is inhibited (Takahashi et al.: Circulation, 88, I-656 (1993)). It is not known that recombinant DNA comprising DNA coding for myosin heavy chain SM1 isoform protein inserted into vector DNA has pharmacological effect and is used for in gene therapy.

For treatment of arteriosclerosis, percutaneous transluminal coronary angioplasty (PTCA) in which a stenosed site is enlarged by a balloon catheter is extensively conducted. The treatment method has the advantages of easier operation and higher degree of success than the bypass surgery etc., while the treatment method has the disadvantage that 30 to 40% of the patients after the treatment suffer from restenosis of blood vessels due to abnormal proliferation of vascular cells. Therefore, there is demand for developments in an agent for treatment of arteriosclerosis effectively used for preventing such restenosis.

›DISCLOSURE OF THE INVENTION · 1 of 2

The present invention includes:

(1) A recombinant DNA comprising DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein inserted into vector DNA.

(2) The recombinant DNA according to (1) wherein the vector DNA is a retrovirus vector, adenovirus vector, adeno-associated virus vector or a plasmid capable of being expressed in an animal.

(3) The recombinant DNA according to (2) wherein the plasmid capable of being expressed in an animal is pCXN2 or PAGE208.

(4) The recombinant DNA according to any one of (1) to (3) wherein the smooth muscle-type myosin heavy chain SM1 isoform protein is of a human smooth muscle type, rabbit muscle type or mouse muscle type.

(5) The recombinant DNA according to any one of (1) to (3) wherein the DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein is the nucleotide sequence shown in SEQ ID NO:1, or the nucleotide sequence shown in SEQ ID NO:1 in which at least one nucleotide is added, deleted or replaced.

(6) DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein, which is the nucleotide sequence shown in SEQ ID NO:1, or the nucleotide sequence shown in SEQ ID NO:1 in which at least one nucleotide is added, deleted or replaced.

(7) A microorganism carrying the recombinant DNA of (5).

(8) The microorganism according to (7) which belongs to the genus Escherichia.

(9) An agent for treatment of arteriosclerosis which comprises the recombinant DNA of any one of (1) to (5).

The DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein includes cDNA or genomic DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein, preferably DNA derived from humans, rabbits, mice, etc.

The nucleotide sequence of the DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein includes the nucleotide sequence of cDNA (SEQ ID NO:5) coding for rabbit SM1 isoform protein (SEQ ID NO:4) (Proc. Natl. Acad. Sci. USA, 88, 10676 (1991)), the nucleotide sequence of the DNA coding for mouse myosin heavy chain SM1 isoform protein as shown in SEQ ID NO:1, and the nucleotide sequence as shown in SEQ ID NO:2 which is specified by homologous regions between the above nucleotide sequences (In the Sequence Listing, Y represents T, U or C; S represents G or C; V represents A, G or C; B represents B, C, T or U; W represents A, T, or U; N represents A, C, G, T, U or a single bond; and the codon YAS (388-390) represents TAC or CAG.). Accordingly, the nucleotide sequence shown in SEQ ID NO:2 contains the nucleotide sequence of SEQ ID NO:1 and the nucleotide sequence of the cDNA coding for rabbit SM1 isoform protein. A partial nucleotide sequence of cDNA coding for human smooth muscle-type myosin heavy chain SM1 isoform protein is also known (Amer. J. of Medical Genetics, 46, 61-67 (1993)), and a nucleotide sequence of this partial nucleotide sequence combined with a partial nucleotide sequence of SEQ ID NO:2 is also contained in the nucleotide sequence of the DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein according to the present invention.

Further, the nucleotide sequence of SEQ ID NO:2 or the combination of a partial nucleotide sequence of SEQ ID NO:2 and the nucleotide sequence of the DNA derived from human smooth muscles, in which one or more nucleotides have been added, deleted or replaced by means of site-directed mutagenesis (Nucleic Acid Research, 10, 6487-6508 (1982)), is also contained in the nucleotide sequence of the DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein according to the present invention.

The vector DNA that can be used includes virus vectors such as retrovirus vector, adenovirus vector and adeno-associated virus vector and plasmids capable of being expressed in an animal such as pCXN2 (Gene, 108, 193-200 (1991)) and PAGE207 (Japanese Patent Laid-Open Publication No. 46841/1994) as well as their modified vectors.

The agent for treatment of arteriosclerosis according to the present invention can be produced by compounding the recombinant DNA comprising DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein inserted into vector DNA as an active ingredient together with a base used in agents for gene therapy. Just before administration, the pharmaceutical preparation can be used in gene therapy for arteriosclerosis, if necessary after encapsulation in liposomes etc. (Proc. Natl. Acad. Sci. USA., 90, 11307 (1993)).

Where the DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein is inserted into a virus vector, a therapeutic agent can be produced by preparing virus particles containing the recombinant DNA and then compounding them together with a base used in agents for gene therapy (Nature Genet., 8, 42(1994)).

The base used in agents for gene therapy may be any base generally used in injections. The base includes, for example, distilled water, a salt solution of sodium chloride, a mixture of sodium chloride and an inorganic salt, or the like, solutions of mannitol, lactose, dextran, glucose, etc., solutions of amino acid such as glycine, arginine, etc., a mixed solution of an organic acid solution or a salt solution and glucose solution, and the like. Further, injections may be prepared in a usual manner as a solution, suspension or dispersion by adding adjuvant such as an osmotic pressure controlling agent, pH adjusting agent, vegetable oils such as sesame oil, soybean oil, etc. or surface active agents such as lecithin, non-ionic surface active agent, etc. to the above base. These injections can also be powdered, lyophilized, etc. to be dissolved just before use.

The agent for treatment of arteriosclerosis can be used as such in the case of a solution, and in the case of a solid it is dissolved in the base previously sterilized if necessary just before use in gene therapy.

The method for administration of the agent for treatment of arteriosclerosis according to the present invention involves topically administrating it into a patients who underwent the PTCA treatment, at a dose of 1 ng to 1 g per day or once after the surgery by means of a catheter etc. such that it can be absorbed into their vascular smooth muscle cells at the target site.

›DISCLOSURE OF THE INVENTION · 2 of 2

The agent for treatment of arteriosclerosis according to the present invention is safe in this dose range.

Hereinafter, the process for producing the recombinant DNA comprising DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein inserted into vector DNA is described.

The recombinant DNA comprising DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein inserted into vector DNA can be obtained according to the usual genetic engineering method described below or its modified method.

A DNA clone coding for smooth muscle-type myosin heavy chain SM1 isoform protein is detected in a cDNA library from tissues composed mainly of smooth muscles in the uterus, aorta, etc., according to the method described by A. Abe et al. (ECL direct DNA labeling detection system manual (Amersham)).

The recombinant DNA comprising the inserted DNA coding for myosin heavy chain SM1 isoform protein can be produced by ligating said DNA fragment to a downstream region from a promoter in a suitable vector DNA (J. Sambrook et al., Molecular Cloning, 2nd Ed., Vol. 1, Cold Spring Harbor Laboratory Press (1989)). Animal cells are used as a host, and a promoter derived from SV40, a promoter from retrovirus, a metallothionein promoter, β-actin promoter etc. can be utilized as the promoter. For the expression, the use of an enhancer is also effective.

The reaction conditions for the above-mentioned recombinant techniques are as follows: The digestion of DNA with a restriction enzyme is carried out by allowing the restriction enzyme in an amount of 0.1 to 100 units, preferably 1 to 3 units per μg DNA to act on 0.1 to 20 μg DNA in a reaction solution usually containing 2 to 200 mM preferably 10 to 40 mM Tris-HCl buffer, pH 6.0 to 9.5, preferably pH 7.0 to 8.0, 0 to 200 mM sodium chloride, and 2 to 20 mM preferably 5 to 10 mM magnesium chloride, at 20 to 70° C. (the optimum temperature is varied depending on the restriction enzyme used) for 15 minutes to 24 hours. The termination of the reaction can be effected usually by heating the reaction solution at 55 to 75° C. for 5 to 30 minutes or by inactivating the restriction enzyme with reagents such as phenol etc. The DNA fragment, or the gapped duplex DNA, generated by digestion with the restriction enzyme can be purified using Prep-A-Gene Matrix (Bio-Rad). The ligation of the DNA fragment can be effected using a DNA ligation kit (Takara Shuzo Co., Ltd.).

The recombinant DNA containing DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein thus constructed in this manner is used to produce a transformant.

The host for the plasmid capable of being expressed in an animal includes, for example, microorganisms belonging to the genus Escherichia, such as Escherichia coli K12·HB101 (H. W. Boyer et al.: J. Mol. Biol.,41, 459 (1969)), DH5 α (D. Hanahan: J. Mol. Biol., 166, 557 (1983)) etc. The transformation of microorganisms of the genus Escherichia can be effected according to the method of Cohen et al. (S. N. Cohen et al.: Proc. Natl. Acad. Sci. USA, 69, 2110 (1972)).

The host used for the virus vector includes animal cells having the ability to produce viruses, such as monkey cell COS-7, Chinese hamster cell CHO, mouse cell BALB/3T3, human cell HeLa, etc.; the host for the retrovirus vector includes ΨCRE, ΨCRIP (Proc. Natl. Acad. Sci. USA, 85, 6460 (1988)), MLV (J. Virol., 65, 1202 (1991)) etc.; and the host for the adenovirus vector and adeno-associated virus vector includes 293 cells derived from human fetal kidney (“jikken Igaku” (Experimental Medicine), 12, 316 (1994)) etc. The introduction of the virus vector into animal cells can be effected using the calcium phosphate method (Virology, 52, 456 (1973)) etc.

The resulting transformant can be cultured in the following manner depending on the difference in the cell species to produce the recombinant DNA.

To culture the transformant from a microorganism of the genus Escherichia as the host, the suitable medium is a liquid medium containing a carbon source, nitrogen source, inorganic substance, etc. necessary for the growth of the transformant. The carbon source includes e.g. glucose, dextrin, soluble starch, sucrose, glycerol, etc. ; the nitrogen source includes e.g. an ammonium salt, peptone, casein, etc.; and the inorganic substance includes e.g. calcium chloride, sodium dihydrogenphosphate, magnesium chloride, etc. A yeast extract, vitamins, etc. may further be added. The pH of the medium is preferably about 5 to 8. To culture the microorganism of the genus Escherichia, the medium is preferably Terrific broth (K. D. Tartof et al.: Bethesda Res. Lab. Focus, 9, 12 (1987)) or the like. The transformant is cultured usually at about 15 to 43° C. for about 8 to 24 hours, if necessary under aeration or stirring. After the culture is finished, the recombinant DNA containing DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein can be obtained through purification by the Birnboim method (Nucleic Acid Res., 7, 1513 (1979)), etc.

To culture the transformant from animal cells as the host, the medium used includes a medium containing about 5 to 20% fetal bovine serum, such as 199 medium (Morgan et al.: Proc. Soci. Biol. Med., 73, 1 (1950)), MEM medium (H. Eagle: Science, 122, 501 (1952)), DMEM (R. Dulbecco et al.: Virology, 8, 396 (1959)) or the like. The pH is preferably in the range of about 6 to 8. The transformant is cultured usually at about 30 to 40° C. for about 18 to 60 hours, if necessary under aeration or stirring.

Because virus particles containing the recombinant DNA are released into the culture supernatant, the recombinant DNA containing DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein can be obtained from the supernatant by concentrating and purifying virus particles by the cesium chloride centrifugation method (Koji Sawada et al.: “Shin Seikagaku Jikken Kouza 2-V” (New Biochemistry Experimental Course 2-V), 33 (1992)), the polyethylene glycol precipitation method (Arch. Virol., 71, 185 (1982)), the filter concentration method (J. Cell. Biol., 111, 217 (1990)), etc.

›BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows a restriction enzyme map of the cDNA coding for mouse smooth muscle-type myosin heavy chain SM1 isoform protein and the location of the coding region.

Plasmid name: SM-1 F text

Plasmid size: 6175 bp

Region in the frame: coding region

A: ApaI

B: BamHI

Sa: SacI

Sm: SmaI

FIG. 2 shows the area of CHO cell colonies selected with hygromycin after introduction of plasmid pSE-SM1-Hyg vs. the area of CHO cell colonies after introduction of plasmid PAGE208.

FIG. 3 shows the area of HeLa cell colonies selected with hygromycin after introduction of plasmid pSE-SM1-Hyg vs. the area of HeLa cell colonies after introduction of plasmid PAGE208.

FIG. 4 shows the comparison among the proliferation rates of the wild type, the PAGE208-introduced clone and the SM1 isoform-expressing clones (SM1-5-2-1, SM1-5-3-3) of CHO cells by the MTT method.

›—◯— CHO

—Δ— PAGE208-1

—— SM1 5-2-1

—▪— SM1 5-3-3

FIG. 5 shows the degrees of thickening of the intima after introduction of the SM1 gene (Example) and β-galactosidase gene (Comparative Example) into the vessel walls in a balloon injured rabbit model.

›BEST MODE FOR CARRYING OUT THE INVENTION

Hereinafter, the present invention is described in more detail with reference to Examples which however are not intended to limit the scope of the present invention.

›Examples4

›EXAMPLE 1 · 1 of 2

Recombinant DNA Comprising DNA Coding for Mouse Smooth Muscle-type Myosin Heavy Chain SM1 Isoform Protein Inserted into Vector DNA

(a) Preparation of a Probe

Plasmids pSMHC29 and pIH61 in which a cDNA fragment coding for rabbit smooth muscle-type myosin heavy chain SM2 isoform protein had been cloned (J. Biol. Chem., 264, 9734-9737 (1989)) were used. These were digested with EcoRI (a product of Takara Shuzo Co., Ltd.; the restriction enzymes used hereinafter are products of Takara Shuzo Co., Ltd. unless otherwise specified) to prepare a cDNA fragment of rabbit smooth muscle-type myosin heavy chain SM2 isoform protein to be inserted. The cDNA fragment was labeled using an ECL direct DNA labeling system (Amersham) to be used as a probe in the following screening.

(b) Screening

A λ gtll vector mouse uterus cDNA library (Clontech) was mixed with E. coli Y1090 (Clontech) and incubated at 37° C. for 15 minutes, followed by adding 0.7% agarose NZY (Nucleic Acid Res., 16, 7583-7600 (1988)), and it was then spread on a 1.5% agar NZY plate. A nylon filter was placed on the plate wherein plaques occurred so that the plaques were transferred to the filter. This filter was subjected to denaturation by alkali treatment and the DNA was fixed by heating at 80° C. for 2 hours. The DNA was hybridized to the probe previously labeled according to the protocol of the ECL direct DNA labeling system in (a) above. Then, the hybridized clones were detected by autoradiography using an ECL detection system (Amersham).

Four clones were identified and each clone was subcloned in pUC119 (Takara Shuzo Co., Ltd.) or pBluescript SK(−) (Stratagene) for analysis of the DNA nucleotide sequence. E. coli DH5α was transformed with the resulting plasmids to give transformants Escherichia coli DH5α/pmsmhc20, pmsmhcC5, pmsmhcN08 and pmsmhcN14. The plasmids produced by the transformants were subjected to stepwise deletion by digestion with an exonuclease or to digestion with a suitable restriction enzyme, followed by self-cyclization or subcloning to prepare template DNA for sequence analysis (Gene, 28, 351-359 (1984)). A fluorescence type DNA sequencer (Applied Biosystems) was used for sequencing and MacMolly (Soft Gene GmbH) was used for data analysis. The nucleotide sequence thus determined is shown in SEQ ID NO:1. The amino acid sequence deduced from the nucleotide sequence is also shown in SEQ ID NO:1.

(d) Construction of an Expression Plasmid for Mouse Smooth Muscle-type Myosin Heavy Chain SM1 Isoform

From the 4 clones previously obtained, plasmid pmSM1 containing cDNA coding for the whole coding region for SM1 isoform was prepared in the following manner (J. Sambrook et al., Molecular Cloning, 2nd Ed., Vol. 1, Cold Spring Harbor Laboratory Press, 1989). To shorten the 5′-non-coding region, plasmid pmsmhcN14 i.e. one of the 4 clones was digested with restriction enzymes BamHI and BglII and a 5-kb DNA fragment was purified and recovered therefrom. The DNA fragment was self-cyclized and transformed into E. coli DH5α to give an ampicillin resistant colony. From the colony, a desired microorganism is picked up and cultured, and plasmid DNA was recovered using a known method (H. C. Birnboim et al.: Nucleic Acid Res., 7, 1513 (1979)). The structure of the plasmid thus obtained was confirmed by agarose gel electrophoresis after digestion with BamHI and KpnI. The plasmid was designated pmsmhcN14′.

The resulting plasmid pmsmhcN14′ was digested with restriction enzymes NsiI (New England Biolabs) and EcoRV, and a 4-kb DNA fragment was purified and recovered therefrom. Separately, plasmid pmsmhcN08 was also digested with NsiI and EcoRV, and a 1-kb DNA fragment was purified, recovered and ligated into the previously obtained 4-kb DNA fragment derived from plasmid pmsmhcN14′, whereby plasmid pmsmhcN14-08 was constructed. The structure of this plasmid was confirmed by digestion with NsiI and EcoRV. The resulting plasmid pmsmhcN14-08 was digested with restriction enzymes ApaI and KpnI and a 5-kb DNA fragment was purified and recovered therefrom. Plasmid pmsmhc20 was also digested in a similar manner with Apal and KpnI, and a 1.5-kb DNA fragment was purified, recovered and ligated into the previously obtained 5-kb DNA fragment, whereby plasmid pmsmhcNl4-08-20 was constructed. The structure of the plasmid was confirmed by digestion with ApaI and KpnI.

The resulting plasmid pmsmhcN14-08-20 was digested with restriction enzyme ApaI and a 6-kb DNA fragment was purified and recovered therefrom. Plasmid pmsmhcC5 was also digested with ApaI, and a 2-kb DNA fragment was purified, recovered and ligated into the 6-kb DNA fragment from pmsmhcN14-08-20 whereby plasmid pmsmhcN14-08-5-20 was constructed. The structure of this plasmid was confirmed by digestion with EcORI and NruI.

Plasmid pmsmhcN14-08-5-20 was digested with restriction enzymes NsiI and NruI and a 8-kb DNA fragment was purified and recovered therefrom. Plasmid pmsmhcN08 was also digested in a similar manner with NsiI and NruI, and a 1.5-kb DNA fragment was purified, recovered and ligated into the previously obtained 8-kb DNA fragment, whereby plasmid pmSM1 containing the whole coding region for mouse smooth muscle-type myosin heavy chain SM1 isoform was constructed. The structure of this plasmid was confirmed by digestion with NsiI and NruI.

The resulting plasmid pmSM1 was digested with restriction enzymes XbaI and KpnI and a 6-kb DNA fragment was purified and recovered therefrom. Separately, expression vector PAGE208 derived from expression vector PAGE207 (Japanese Patent Laid-Open Publication No. 46841/1994) by deleting an SmaI site located in a promoter region for hygromycin B resistance gene in PAGE207 was digested with XbaI and KpnI, and a 6-kb DNA fragment was purified, recovered, and ligated into the previously obtained 6-kb DNA fragment derived from plasmid pmSM1, whereby expression plasmid pSE-SM1-Hyg was constructed. The structure of the plasmid was confirmed by digestion with XbaI and KpnI.

›EXAMPLE 1 · 2 of 2

A DNA fragment obtained by digestion of plasmid pmSM1 with XbaI and KpnI was blunt-ended with a DNA blunting kit (Takara Shuzo Co., Ltd.). Separately, expression vector pCXN2 was digested with XhoI and blunt-ended with the DNA blunting kit (Takara Shuzo Co., Ltd.). These 2 kinds of DNA were ligated to construct expression plasmid pCAG-SM1. The structure of the plasmid was confirmed by digestion with BglII.

The above plasmid pSE-SM1-Hyg was inserted into Escherichia coli whereby a transformant was obtained.

The transformant has been deposited as Escherichia coli pSE-SM1-Hyg (FERM BP-4974) with the National Institute of Bioscience and Human-Technology, Agency of Industrial Science and Technology, 1-3, Higashi 1 chome, Tsukuba-shi, Ibaraki-ken, Japan, since Jan. 24, 1995.

›EXAMPLE 2

Effect of Expression of SM1 Isoform in CHO Cells and HeLa Cells on Cell Proliferation

2 μg of expression plasmid pSE-SM1-Hyg and 4 μl of lipofectamine™ (Gibco) were mixed and incubated for 15 minutes to produce a complex. The complex was added to CHO cells or HeLa cells and incubated at 37° C. for 5 hours so that the gene was introduced into the cells. The cells were diluted and spread on a vessel, and cells into which the gene had been introduced were selected in the presence of hygromycine™ (Sigma). About 10 days later, the drug resistant cell colonies were stained with Coomassie Blue™ (Bio-Rad) and their area was measured with an image analyzer. As a result, many of the colonies had smaller areas than those of the colonies carrying the introduced control vector PAGE208, indicating that their proliferation was inhibited by the expression of the SM1 isoform (FIGS. 2 and 3 ). Separately, the hygromycin resistant CHO cells were cloned and the resulting clones SM1-5-2-1 and SM1-5-3-3 were examined for their proliferation rate. The result indicated that their proliferation rate was lower than that of the wild-type or PAGE208-introduced cells (FIG. 4 ).

›EXAMPLE 3

Inhibitory Effect of Forcible Introduction of SM1 Gene into Vessel Walls in a Balloon Injured Rabbit Model on Thickening of the Intima

A PTCA catheter at 3 French (Fr) was inserted under X-raying through the femoral artery into the right common carotid artery of a Japanese white domestic rabbit weighing 2.4 to 3.2 kg. The right common carotid artery was abraded 3 times with a balloon expanded at 10 atm. to injure the vessel wall. Three days after the abrasion, the injured portion of the vessel was infused with a mixture of 300 mg pCAG-SM1 and lipofectin (Gibco BRL) through a Wolinsky type infusion catheter (a product of BARD) at 6 atm. For Comparative Example, other domestic rabbits were treated in the same manner, and in place of pCAG-SM1, an expression vector for β-galactosidase was introduced into the injured portion of the vessel.

Three days after the infusion, the animals were sacrificed by perfusion with heparin physiological saline and their right common carotid arteries were removed and cut thin, followed by adding 1 ml ISOGEN (Wako Pure Chemical Industries, Ltd.), and the tissues were homogenized with a Polytron homogenizer. The whole RNA was extracted by the AGPC method (P. Chomczynski and N. Sacchi: Anal. Biochem., 162, 156-159 (1987)), and the MRNA for SM1 was amplified by the RT-PCR method. The amplified DNAs from the Example (introduction of SM1 gene) and Comparative Example (introduction of β-galactosidase gene) and their fragments digested with restriction enzyme NheI were subjected to agarose gel electrophoresis. The SM1 band was detected strongly in the Example and faintly in the Comparative Example, and the band in only the Example was digested with NheI. The supported that the SM1 gene derived from the mouse was expressed in the Example because the digestion site of the restriction enzyme is not present in the DNA for SM1 derived from the domestic rabbit but present in the mouse SM1.

Two weeks after introduction of the gene, the rabbits were sacrificed by perfusion with PBS containing 2% paraformaldehyde and fixed, and their right common carotid arteries were removed. The part into which the infusion catheter had been introduced was dissected out and embedded in paraffin, and a thin round section of the vessel was prepared. One vessel was divided equally into 4 tissues and 1 section was prepared from each tissue and stained with hematoxylineosin. The vessel was divided along the internal elastic plate as the boundary into the intima and media, and the area of each was determined. The intima area/media area ratio was determined for each section and the mean was determined. The degree of thickening of the intima was determined using the mean. From the degree of thickening of the intima as determined for 3 animals in the Example and 3 animals in the Comparative Example, it was confirmed that thickening of the intima was inhibited by forcible introduction of the SM1 gene (FIG. 5 ).

The results suggested that plasmids pSE-SM1-Hyg and pCAG-SM1 possess the effect of depressing cell proliferation and are effective in restenosis of arteriosclerosis, caused by abnormal proliferation of vascular cells.

›INDUSTRIAL APPLICABILITY · 1 of 5

It was revealed that the proliferation of animal cells is inhibited when the recombinant DNA comprising DNA coding for smooth muscle-type myosin heavy chain SM1 isoform protein inserted into vector DNA according to the present invention is expressed in the animal cells. The effect was also observed in human-derived HeLa cells. Therefore, the recombinant DNA of the present invention can be used effectively as an agent for gene therapy of restenosis after PTCA treatment.

agatttggac catcccagcc tgggatcagt gccagatccg agctctccat ccggtgtcct 60

cctgctaatc cacccccgga gtagatctgg gaccaccaga catc atg gcg cag aaa 116

Met Ala Gln Lys

ggg cag ctc agc gat gat gag aag ttc ctc ttt gtg gat aaa aac ttc 164

Gly Gln Leu Ser Asp Asp Glu Lys Phe Leu Phe Val Asp Lys Asn Phe

5 10 15 20

atg aac agc cca atg gct cag gcc gac tgg gta gcc aag aag ctg gtg 212

Met Asn Ser Pro Met Ala Gln Ala Asp Trp Val Ala Lys Lys Leu Val

25 30 35

tgg gtc cct tca gag aag cag ggc ttc gaa gca gcc agc atc aag gag 260

Trp Val Pro Ser Glu Lys Gln Gly Phe Glu Ala Ala Ser Ile Lys Glu

40 45 50

gag aag ggc gat gag gtg gtc gtg gag ttg gtg gaa aat gga aag aag 308

Glu Lys Gly Asp Glu Val Val Val Glu Leu Val Glu Asn Gly Lys Lys

55 60 65

gtc aca gtt ggc aaa gat gac atc caa aaa atg aac cca ccc aag ttc 356

Val Thr Val Gly Lys Asp Asp Ile Gln Lys Met Asn Pro Pro Lys Phe

70 75 80

tct aag gtg gag gac atg gca gag ctg acg tgc ctc aat gag gcc tct 404

Ser Lys Val Glu Asp Met Ala Glu Leu Thr Cys Leu Asn Glu Ala Ser

85 90 95 100

gtg ctg cac aac ctg agg gag cga tac ttc tca ggc ctc atc tat acc 452

Val Leu His Asn Leu Arg Glu Arg Tyr Phe Ser Gly Leu Ile Tyr Thr

105 110 115

tac tct ggc ctc ttc tgt gtg gtg gtc aac ccc tac aag tac cta ccc 500

Tyr Ser Gly Leu Phe Cys Val Val Val Asn Pro Tyr Lys Tyr Leu Pro

120 125 130

atc tac tca gaa aag atc gtg gat atg tac aag ggc aag aag agg cat 548

Ile Tyr Ser Glu Lys Ile Val Asp Met Tyr Lys Gly Lys Lys Arg His

135 140 145

gag atg ccg cct cac atc tat gcc att gcc gac aca gcc tac aga agc 596

Glu Met Pro Pro His Ile Tyr Ala Ile Ala Asp Thr Ala Tyr Arg Ser

150 155 160

atg cta caa gat cgt gaa gac cag tcc att ctg tgc aca ggt gag tct 644

Met Leu Gln Asp Arg Glu Asp Gln Ser Ile Leu Cys Thr Gly Glu Ser

165 170 175 180

gga gcc gga aag aca gag aac aca cag aaa gtc ata cag tac ttg gct 692

Gly Ala Gly Lys Thr Glu Asn Thr Gln Lys Val Ile Gln Tyr Leu Ala

185 190 195

gtg gtg gcg tcc tcc cac aag ggc aag aaa gac agc agc atc acg ggg 740

Val Val Ala Ser Ser His Lys Gly Lys Lys Asp Ser Ser Ile Thr Gly

200 205 210

gag ctg gaa aag cag ctt cta cag gca aac cca atc ctg gag gct ttc 788

Glu Leu Glu Lys Gln Leu Leu Gln Ala Asn Pro Ile Leu Glu Ala Phe

215 220 225

ggc aat gcg aaa acc gtc aag aac gac aac tcc tct cgc ttt ggc aag 836

Gly Asn Ala Lys Thr Val Lys Asn Asp Asn Ser Ser Arg Phe Gly Lys

230 235 240

ttc att cgc atc aac ttc gat gtc act ggt tac att gta ggt gcc aat 884

Phe Ile Arg Ile Asn Phe Asp Val Thr Gly Tyr Ile Val Gly Ala Asn

245 250 255 260

att gaa aca tat ctt ctg gaa aag tct agg gct att cga cag gct agg 932

Ile Glu Thr Tyr Leu Leu Glu Lys Ser Arg Ala Ile Arg Gln Ala Arg

265 270 275

gat gag aga aca ttt cac atc ttc tac tac ctg ctc gcc gga gcc aag 980

Asp Glu Arg Thr Phe His Ile Phe Tyr Tyr Leu Leu Ala Gly Ala Lys

280 285 290

gaa aag atg aaa agt gac ctg ctt ttg gag agc ttc aac agc tac aca 1028

Glu Lys Met Lys Ser Asp Leu Leu Leu Glu Ser Phe Asn Ser Tyr Thr

295 300 305

ttt tta tcc aat ggc ttt gtg ccc atc cca gct gca caa gat gat gag 1076

Phe Leu Ser Asn Gly Phe Val Pro Ile Pro Ala Ala Gln Asp Asp Glu

310 315 320

atg ttc cag gag aca ctg gaa gcc atg tct atc atg ggc ttc aat gaa 1124

Met Phe Gln Glu Thr Leu Glu Ala Met Ser Ile Met Gly Phe Asn Glu

325 330 335 340

gag gaa cag cta gcc atc ttg aag gta gta tca tct gtc ctt cag ctt 1172

Glu Glu Gln Leu Ala Ile Leu Lys Val Val Ser Ser Val Leu Gln Leu

345 350 355

gga aac att gtc ttc aag aag gag cga aac aca gac cag gca tcc atg 1220

Gly Asn Ile Val Phe Lys Lys Glu Arg Asn Thr Asp Gln Ala Ser Met

360 365 370

cct gat aac aca gcg gct cag aaa gtt tgc cac ctc gtg ggg att aat 1268

Pro Asp Asn Thr Ala Ala Gln Lys Val Cys His Leu Val Gly Ile Asn

375 380 385

gtg aca gat ttc act aga gcc atc ctg acc cca cgt atc aaa gtt gga 1316

Val Thr Asp Phe Thr Arg Ala Ile Leu Thr Pro Arg Ile Lys Val Gly

390 395 400

cgg gat gtg gtg cag aag gct cag acc aaa gaa cag gct gac ttc gcc 1364

Arg Asp Val Val Gln Lys Ala Gln Thr Lys Glu Gln Ala Asp Phe Ala

405 410 415 420

atc gag gcc tta gcc aag gcc acc tat gag cgc ctt ttc cga tgg att 1412

Ile Glu Ala Leu Ala Lys Ala Thr Tyr Glu Arg Leu Phe Arg Trp Ile

425 430 435

ctc agc cgt gta aac aag gcc ttg gac aag acc cat cgg cag ggg gcc 1460

Leu Ser Arg Val Asn Lys Ala Leu Asp Lys Thr His Arg Gln Gly Ala

440 445 450

tcc ttc ctg ggc att ctg gat att gct ggg ttt gaa atc ttt gag gta 1508

Ser Phe Leu Gly Ile Leu Asp Ile Ala Gly Phe Glu Ile Phe Glu Val

455 460 465

aac tcc ttc gag cag ctg tgc atc aac tac acc aac gag aag ctg cag 1556

Asn Ser Phe Glu Gln Leu Cys Ile Asn Tyr Thr Asn Glu Lys Leu Gln

470 475 480

cag ctg ttc aac cac acg atg ttc atc ctg gag cag gaa gag tac cag 1604

Gln Leu Phe Asn His Thr Met Phe Ile Leu Glu Gln Glu Glu Tyr Gln

485 490 495 500

cga gag ggc atc gag tgg aac ttc atc gac ttc ggc ctg gac ctg cag 1652

Arg Glu Gly Ile Glu Trp Asn Phe Ile Asp Phe Gly Leu Asp Leu Gln

505 510 515

cct agt att gag ctg att gag cgg ccg aac aac cct ccc ggt gtg ctg 1700

Pro Ser Ile Glu Leu Ile Glu Arg Pro Asn Asn Pro Pro Gly Val Leu

520 525 530

gcc ctg ctg gat gaa gaa tgc tgg ttc ccc aaa gct aca gac aag tct 1748

Ala Leu Leu Asp Glu Glu Cys Trp Phe Pro Lys Ala Thr Asp Lys Ser

535 540 545

ttt gtg gag aag cta tgc tca gag cag gga aat cac ccc aaa ttt cag 1796

Phe Val Glu Lys Leu Cys Ser Glu Gln Gly Asn His Pro Lys Phe Gln

550 555 560

aag ccc aag cag cta aag gac aaa aca gag ttc tcc atc atc cac tat 1844

Lys Pro Lys Gln Leu Lys Asp Lys Thr Glu Phe Ser Ile Ile His Tyr

565 570 575 580

gct ggg aag gtg gac tac aat gca agt gcc tgg ctg acc aag aac atg 1892

Ala Gly Lys Val Asp Tyr Asn Ala Ser Ala Trp Leu Thr Lys Asn Met

585 590 595

gac ccg cta aat gac aat gtg aca tca ctc ctc aat gcc tcc tct gac 1940

Asp Pro Leu Asn Asp Asn Val Thr Ser Leu Leu Asn Ala Ser Ser Asp

›INDUSTRIAL APPLICABILITY · 2 of 5

600 605 610

aag ttt gtg gct gac ctg tgg aag gat gtg gac cgc att gtg ggg ctg 1988

Lys Phe Val Ala Asp Leu Trp Lys Asp Val Asp Arg Ile Val Gly Leu

615 620 625

gac cag atg gcc aag atg act gag agc tca ctg ccc agt gcc tca aag 2036

Asp Gln Met Ala Lys Met Thr Glu Ser Ser Leu Pro Ser Ala Ser Lys

630 635 640

acc aaa aag ggc atg ttc cgc acc gtg gga cag ctc tac aaa gag cag 2084

Thr Lys Lys Gly Met Phe Arg Thr Val Gly Gln Leu Tyr Lys Glu Gln

645 650 655 660

ttg ggg aaa ctg atg gct aca ctg cgc aat acc acg gct aac ttt gtg 2132

Leu Gly Lys Leu Met Ala Thr Leu Arg Asn Thr Thr Ala Asn Phe Val

665 670 675

cgc tgc atc atc ccc aac cat gag aag agg tct ggc aag ctg gat gca 2180

Arg Cys Ile Ile Pro Asn His Glu Lys Arg Ser Gly Lys Leu Asp Ala

680 685 690

ttt cta gtg ctg gaa cag ctg cgc tgc aac ggt gtg ttg gaa ggc atc 2228

Phe Leu Val Leu Glu Gln Leu Arg Cys Asn Gly Val Leu Glu Gly Ile

695 700 705

cgc atc tgc cgt cag ggc ttc ccc aac agg att gtc ttc caa gag ttc 2276

Arg Ile Cys Arg Gln Gly Phe Pro Asn Arg Ile Val Phe Gln Glu Phe

710 715 720

cgg caa cgc tat gag atc ctg gca gcg aac gcc atc ccc aaa ggc ttc 2324

Arg Gln Arg Tyr Glu Ile Leu Ala Ala Asn Ala Ile Pro Lys Gly Phe

725 730 735 740

atg gat gga aag caa gcc tgc att ctc atg atc aaa gcc ctc gaa ctt 2372

Met Asp Gly Lys Gln Ala Cys Ile Leu Met Ile Lys Ala Leu Glu Leu

745 750 755

gac cct aac ctg tac agg att ggg cag agc aaa atc ttc ttc cga acg 2420

Asp Pro Asn Leu Tyr Arg Ile Gly Gln Ser Lys Ile Phe Phe Arg Thr

760 765 770

ggg gtc ctg gcc cac cta gag gag gaa cga gac ctg aaa att act gat 2468

Gly Val Leu Ala His Leu Glu Glu Glu Arg Asp Leu Lys Ile Thr Asp

775 780 785

gtc atc atg gcc ttc cag gca atg tgt cgt ggc tac ctt gcc aga aag 2516

Val Ile Met Ala Phe Gln Ala Met Cys Arg Gly Tyr Leu Ala Arg Lys

790 795 800

gcc ttc acc aag agg cag caa cag ctg aca gcc atg aag gtg atc cag 2564

Ala Phe Thr Lys Arg Gln Gln Gln Leu Thr Ala Met Lys Val Ile Gln

805 810 815 820

agg aac tgc gct gcc tac ctt aag ctc cgc aac tgg caa tgg tgg cgg 2612

Arg Asn Cys Ala Ala Tyr Leu Lys Leu Arg Asn Trp Gln Trp Trp Arg

825 830 835

ctc ttc acc aaa gta aag cca ttg ctc cag gtg aca cgg cag gag gag 2660

Leu Phe Thr Lys Val Lys Pro Leu Leu Gln Val Thr Arg Gln Glu Glu

840 845 850

gag atg cag gcc aag gag gag gag atg caa aag atc acg gag cgg cag 2708

Glu Met Gln Ala Lys Glu Glu Glu Met Gln Lys Ile Thr Glu Arg Gln

855 860 865

cag aag gca gag act gag ttg aag gag ctg gag cag aag cac act cag 2756

Gln Lys Ala Glu Thr Glu Leu Lys Glu Leu Glu Gln Lys His Thr Gln

870 875 880

ctg gct gag gag aag act ctg ctg cag gag cag ttg cag gca gag aca 2804

Leu Ala Glu Glu Lys Thr Leu Leu Gln Glu Gln Leu Gln Ala Glu Thr

885 890 895 900

gag ctg tat gct gag tct gag gag atg cgg gtc cgg ttg gca gcc aag 2852

Glu Leu Tyr Ala Glu Ser Glu Glu Met Arg Val Arg Leu Ala Ala Lys

905 910 915

aag cag gaa ctg gag gag atc cta cat gag atg gag gcc cgc ctg gag 2900

Lys Gln Glu Leu Glu Glu Ile Leu His Glu Met Glu Ala Arg Leu Glu

920 925 930

gaa gag gaa gac cgg cgc cag caa cta cag gct gag agg aag aag atg 2948

Glu Glu Glu Asp Arg Arg Gln Gln Leu Gln Ala Glu Arg Lys Lys Met

935 940 945

gct cag cag atg cta gac ctg gag gag caa ctg gag gag gaa gag gcc 2996

Ala Gln Gln Met Leu Asp Leu Glu Glu Gln Leu Glu Glu Glu Glu Ala

950 955 960

gcc aga cag aaa cta cag cta gag aag gtc acg gct gag gcc aag atc 3044

Ala Arg Gln Lys Leu Gln Leu Glu Lys Val Thr Ala Glu Ala Lys Ile

965 970 975 980

aag aaa ctg gag gat gac atc ttg gtt atg gat gat cag aac agt aaa 3092

Lys Lys Leu Glu Asp Asp Ile Leu Val Met Asp Asp Gln Asn Ser Lys

985 990 995

ctt tca aaa gaa cga aaa ctc ctt gaa gag agg gtc agc gac ttg aca 3140

Leu Ser Lys Glu Arg Lys Leu Leu Glu Glu Arg Val Ser Asp Leu Thr

1000 1005 1010

acc aac cta gca gaa gag gaa gaa aag gct aaa aac ctc aca aag ctg 3188

Thr Asn Leu Ala Glu Glu Glu Glu Lys Ala Lys Asn Leu Thr Lys Leu

1015 1020 1025

aag agc aag cat gag tct atg atc tca gag ctg gag gtg agg ctg aag 3236

Lys Ser Lys His Glu Ser Met Ile Ser Glu Leu Glu Val Arg Leu Lys

1030 1035 1040

aaa gag gag aag agc cgg cag gag ctg gag aaa ctc aag agg aaa ctg 3284

Lys Glu Glu Lys Ser Arg Gln Glu Leu Glu Lys Leu Lys Arg Lys Leu

1045 1050 1055 1060

gag ggt gat gcc agt gac ttc cat gag cag atc gct gac ttg cag gcc 3332

Glu Gly Asp Ala Ser Asp Phe His Glu Gln Ile Ala Asp Leu Gln Ala

1065 1070 1075

cag att gca gag ctc aag atg cag ctg gca aag aaa gag gaa gag cta 3380

Gln Ile Ala Glu Leu Lys Met Gln Leu Ala Lys Lys Glu Glu Glu Leu

1080 1085 1090

cag gca gct cta gcc agg ctt gat gaa gag atc gcc cag aaa aac aat 3428

Gln Ala Ala Leu Ala Arg Leu Asp Glu Glu Ile Ala Gln Lys Asn Asn

1095 1100 1105

gcc cta aag aag att cgc gag cta gag ggc cat atc tca gac cta caa 3476

Ala Leu Lys Lys Ile Arg Glu Leu Glu Gly His Ile Ser Asp Leu Gln

1110 1115 1120

gag gac cta gac tca gag cgg gct gcc agg aac aag gcc gag aag cag 3524

Glu Asp Leu Asp Ser Glu Arg Ala Ala Arg Asn Lys Ala Glu Lys Gln

1125 1130 1135 1140

aag cga gac ctg ggg gag gag ctg gag gca ctc aag acg gag ctg gag 3572

Lys Arg Asp Leu Gly Glu Glu Leu Glu Ala Leu Lys Thr Glu Leu Glu

1145 1150 1155

gat acg ctg gac agc aca gct acc cag cag gag ctc aga gcc aag agg 3620

Asp Thr Leu Asp Ser Thr Ala Thr Gln Gln Glu Leu Arg Ala Lys Arg

1160 1165 1170

gaa cag gag gtg aca gtg ctg aag aag gcc ctg gat gag gag acg cgg 3668

Glu Gln Glu Val Thr Val Leu Lys Lys Ala Leu Asp Glu Glu Thr Arg

1175 1180 1185

tcc cat gag gcc cag gtc cag gag atg agg cag aag cac aca cag gca 3716

Ser His Glu Ala Gln Val Gln Glu Met Arg Gln Lys His Thr Gln Ala

1190 1195 1200

gtg gag gaa ctc aca gag cag ctg gag cag ttc aaa agg gcc aag gca 3764

Val Glu Glu Leu Thr Glu Gln Leu Glu Gln Phe Lys Arg Ala Lys Ala

1205 1210 1215 1220

aac ctg gac aaa agc aag cag aca ctg gag aag gag aac gcg gac ctg 3812

Asn Leu Asp Lys Ser Lys Gln Thr Leu Glu Lys Glu Asn Ala Asp Leu

1225 1230 1235

gct ggg gag ctg cgt gtc ctg ggc cag gcg aag cag gag gtg gaa cac 3860

Ala Gly Glu Leu Arg Val Leu Gly Gln Ala Lys Gln Glu Val Glu His

1240 1245 1250

aag aag aag aag ctg gag gtg cag ctg cag gat ctg cag tcc aag tgc 3908

Lys Lys Lys Lys Leu Glu Val Gln Leu Gln Asp Leu Gln Ser Lys Cys

1255 1260 1265

agt gat ggg gag cgt gcc cgg gct gag ctc agt gac aag gtc cac aag 3956

Ser Asp Gly Glu Arg Ala Arg Ala Glu Leu Ser Asp Lys Val His Lys

›INDUSTRIAL APPLICABILITY · 3 of 5

1270 1275 1280

cta cag aat gaa gtg gag agt gtc act ggc atg ctc aat gag gca gag 4004

Leu Gln Asn Glu Val Glu Ser Val Thr Gly Met Leu Asn Glu Ala Glu

1285 1290 1295 1300

ggc aaa gcc atc aaa ctg gcc aaa gat gtg gct tcc ctt gga tcc cag 4052

Gly Lys Ala Ile Lys Leu Ala Lys Asp Val Ala Ser Leu Gly Ser Gln

1305 1310 1315

ctt cag gac acc caa gag ctg ctc caa gaa gaa acc cgg cag aag ctc 4100

Leu Gln Asp Thr Gln Glu Leu Leu Gln Glu Glu Thr Arg Gln Lys Leu

1320 1325 1330

aat gtg tct acc aag ctg cgt cag ttg gaa gat gaa agg aac agc ctg 4148

Asn Val Ser Thr Lys Leu Arg Gln Leu Glu Asp Glu Arg Asn Ser Leu

1335 1340 1345

cag gac cag ctg gat gag gag atg gag gct aag caa aac ctg gag cgc 4196

Gln Asp Gln Leu Asp Glu Glu Met Glu Ala Lys Gln Asn Leu Glu Arg

1350 1355 1360

cat gtc tca aca ctg aac att cag ctc tca gac tct aag aag aag ctg 4244

His Val Ser Thr Leu Asn Ile Gln Leu Ser Asp Ser Lys Lys Lys Leu

1365 1370 1375 1380

cag gac ttt gca agt acc atc gag gtc atg gag gag ggg aag aag agg 4292

Gln Asp Phe Ala Ser Thr Ile Glu Val Met Glu Glu Gly Lys Lys Arg

1385 1390 1395

tta cag aaa gag atg gag ggc ctc agc cag cag tat gag gag aag gcg 4340

Leu Gln Lys Glu Met Glu Gly Leu Ser Gln Gln Tyr Glu Glu Lys Ala

1400 1405 1410

gct gcc tat gac aaa ctg gag aaa acc aag aac agg ctc cag cag gag 4388

Ala Ala Tyr Asp Lys Leu Glu Lys Thr Lys Asn Arg Leu Gln Gln Glu

1415 1420 1425

ctg gat gac ctg gtc gtg gac ttg gac aac cag cgg caa ctg gta tcc 4436

Leu Asp Asp Leu Val Val Asp Leu Asp Asn Gln Arg Gln Leu Val Ser

1430 1435 1440

aat ctg gaa aag aag cag aag aaa ttt gac cag ttg tta gct gag gag 4484

Asn Leu Glu Lys Lys Gln Lys Lys Phe Asp Gln Leu Leu Ala Glu Glu

1445 1450 1455 1460

aag aac atc tcc tcc aag tat gcg gat gag aga gac cga gct gaa gca 4532

Lys Asn Ile Ser Ser Lys Tyr Ala Asp Glu Arg Asp Arg Ala Glu Ala

1465 1470 1475

gag gcc agg gaa aag gag aca aag gct ttg tct cta gcc cgg gcc ctg 4580

Glu Ala Arg Glu Lys Glu Thr Lys Ala Leu Ser Leu Ala Arg Ala Leu

1480 1485 1490

gag gaa gcc ctg gaa gcc aaa gaa gag ctg gag agg acc aac aag atg 4628

Glu Glu Ala Leu Glu Ala Lys Glu Glu Leu Glu Arg Thr Asn Lys Met

1495 1500 1505

ctc aaa gct gag atg gaa gac ctg gtc agc tcc aag gat gat gta ggc 4676

Leu Lys Ala Glu Met Glu Asp Leu Val Ser Ser Lys Asp Asp Val Gly

1510 1515 1520

aag aac gtg cat gaa ctg gag aag tcc aag cgt gcc ttg gag acc cag 4724

Lys Asn Val His Glu Leu Glu Lys Ser Lys Arg Ala Leu Glu Thr Gln

1525 1530 1535 1540

atg gaa gag atg aaa acc cag ctg gag gag tcg gag gat gac gtg cag 4772

Met Glu Glu Met Lys Thr Gln Leu Glu Glu Ser Glu Asp Asp Val Gln

1545 1550 1555

gcc act gag gat gcc aag ctg cgg cta gag gtc aac atg cag gcc ctc 4820

Ala Thr Glu Asp Ala Lys Leu Arg Leu Glu Val Asn Met Gln Ala Leu

1560 1565 1570

aag ggc cag ttt gaa cgc gat ctc cag gct cgg gat gaa cag aat gag 4868

Lys Gly Gln Phe Glu Arg Asp Leu Gln Ala Arg Asp Glu Gln Asn Glu

1575 1580 1585

gag aag agg agg cag cta cag cgg cag ctg cac gag tat gag aca gaa 4916

Glu Lys Arg Arg Gln Leu Gln Arg Gln Leu His Glu Tyr Glu Thr Glu

1590 1595 1600

ctg gaa gat gaa cgg aag cag aga gct ctg gcg gcg gca gct aag aag 4964

Leu Glu Asp Glu Arg Lys Gln Arg Ala Leu Ala Ala Ala Ala Lys Lys

1605 1610 1615 1620

aag ctg gaa ggg gac cta aaa gac cta gag ctc cag gct gac tca gcc 5012

Lys Leu Glu Gly Asp Leu Lys Asp Leu Glu Leu Gln Ala Asp Ser Ala

1625 1630 1635

atc aaa ggg agg gag gaa gcc atc aag cag ctt cga aaa ctg cag gct 5060

Ile Lys Gly Arg Glu Glu Ala Ile Lys Gln Leu Arg Lys Leu Gln Ala

1640 1645 1650

cag atg aag gac ttc caa aga gag ctg gat gat gcc cgt gcc tcc agg 5108

Gln Met Lys Asp Phe Gln Arg Glu Leu Asp Asp Ala Arg Ala Ser Arg

1655 1660 1665

gat gag atc ttt gcc acc tca aaa gag aat gag aag aaa gcc aag agt 5156

Asp Glu Ile Phe Ala Thr Ser Lys Glu Asn Glu Lys Lys Ala Lys Ser

1670 1675 1680

ctg gag gca gac ctc atg cag ctc caa gag gac ctg gca gca gct gag 5204

Leu Glu Ala Asp Leu Met Gln Leu Gln Glu Asp Leu Ala Ala Ala Glu

1685 1690 1695 1700

aga gct cgc aag caa gct gac ctg gag aag gag gag ctg gcc gag gag 5252

Arg Ala Arg Lys Gln Ala Asp Leu Glu Lys Glu Glu Leu Ala Glu Glu

1705 1710 1715

ctg gct agc agc ttg tca gga agg aat aca ctg cag gat gag aag cgc 5300

Leu Ala Ser Ser Leu Ser Gly Arg Asn Thr Leu Gln Asp Glu Lys Arg

1720 1725 1730

cgc ctg gag gca agg atc gcc caa cta gag gag gag ctg gag gaa gag 5348

Arg Leu Glu Ala Arg Ile Ala Gln Leu Glu Glu Glu Leu Glu Glu Glu

1735 1740 1745

cag ggc aac atg gag gcc atg agt gat aga gta cgc aag gcc aca ctg 5396

Gln Gly Asn Met Glu Ala Met Ser Asp Arg Val Arg Lys Ala Thr Leu

1750 1755 1760

cag gct gag caa ctg agc aat gag ctg gcc aca gaa cgc agc acg gct 5444

Gln Ala Glu Gln Leu Ser Asn Glu Leu Ala Thr Glu Arg Ser Thr Ala

1765 1770 1775 1780

cag aag aat gag agc gca cgg caa cag ctg gag cgc cag aac aag gaa 5492

Gln Lys Asn Glu Ser Ala Arg Gln Gln Leu Glu Arg Gln Asn Lys Glu

1785 1790 1795

ctg cga agc aag ttg cag gag gta gaa ggt gct gtc aaa gcc aag ctc 5540

Leu Arg Ser Lys Leu Gln Glu Val Glu Gly Ala Val Lys Ala Lys Leu

1800 1805 1810

aag tcc act gtt gcg gcg ctg gag gcc aag att gca cag ctg gag gag 5588

Lys Ser Thr Val Ala Ala Leu Glu Ala Lys Ile Ala Gln Leu Glu Glu

1815 1820 1825

cag gtt gaa cag gag gcc aga gag aaa cag gcg gcc acc aag tcg ctg 5636

Gln Val Glu Gln Glu Ala Arg Glu Lys Gln Ala Ala Thr Lys Ser Leu

1830 1835 1840

aag caa aag gac aag aag cta aag gag gtc ctg ctg cag gtg gag gat 5684

Lys Gln Lys Asp Lys Lys Leu Lys Glu Val Leu Leu Gln Val Glu Asp

1845 1850 1855 1860

gag cgc aag atg gca gag cag tac aag gag cag gca gag aaa gga aac 5732

Glu Arg Lys Met Ala Glu Gln Tyr Lys Glu Gln Ala Glu Lys Gly Asn

1865 1870 1875

acc aag gtc aag cag ctg aag agg cag ctg gaa gag gca gag gag gag 5780

Thr Lys Val Lys Gln Leu Lys Arg Gln Leu Glu Glu Ala Glu Glu Glu

1880 1885 1890

tcc cag tgc atc aac gcc aac cgc agg aag ctg cag cgg gag cta gat 5828

Ser Gln Cys Ile Asn Ala Asn Arg Arg Lys Leu Gln Arg Glu Leu Asp

1895 1900 1905

gag gcc aca gag agc aat gag gcc atg ggc cgt gag gtg aac gcc ctc 5876

Glu Ala Thr Glu Ser Asn Glu Ala Met Gly Arg Glu Val Asn Ala Leu

1910 1915 1920

aag agc aaa ctc agg aga gga aac gag gct tca ttt gtt cct tcc aga 5924

Lys Ser Lys Leu Arg Arg Gly Asn Glu Ala Ser Phe Val Pro Ser Arg

1925 1930 1935 1940

›INDUSTRIAL APPLICABILITY · 4 of 5

agg gct ggg ggc cgt aga gtt att gaa aac aca gat ggt tct gaa gaa 5972

Arg Ala Gly Gly Arg Arg Val Ile Glu Asn Thr Asp Gly Ser Glu Glu

1945 1950 1955

gaa atg gac gct cgg gac tca gac ttc aat gga acc aaa gcc agt gaa 6020

Glu Met Asp Ala Arg Asp Ser Asp Phe Asn Gly Thr Lys Ala Ser Glu

1960 1965 1970

taaattcagg attggacacc atgtcaggga aaacagaaca ctaaacgaca gcagagccca 6080

gcagactgct tagcacttgt gtccattcgt tctcaagtca cagaaatcac tccacccctc 6140

accaggagtc aaccacagcc ctgcacaaag ggtgt 6175

atggcgcaga anggncanct cagcgangat gagaagttcc tctttgtgga naanaacttc 60

atnaacagcc cnntggcnca ggccgactgg gtngccaaga ngctggtgtg ggtcccttcn 120

gagaagcagg gcttcgangc ngccagcatc aaggaggaga agggngatga ggtggtcgtg 180

gagntggtgg anaatggnaa gaaggtcacn gtnggcaang atgacatcca naanatgaac 240

ccncccaagt tctcnaaggt ggangacatg gcngagctga cgtgbctcaa ngangcntcn 300

gtgctgcaca acctgaggga gvgntacttc tcnggnctca tctanacnta ctcnggcctc 360

ttctgbgtgg tggtcaaccc ctacaagyas ctncccatct actcnganaa gatcgtggan 420

atgtacaagg gcaagaagag gcangagatg ccnccncaca tctabgccat ngccgacacn 480

gcctacagna gcatgctnca ngatcgngan gaccagtcca ttctntgcac aggngagtct 540

ggagccggna agacngagaa cacnvagaaa gtcatncagt acntggcngt ggtggcntcn 600

tcccacaagg gcaagaanga cannagcatc acgggggagc tgganaagca gcttctncan 660

gcaaacccna tcctggaggc nttnggcaan gcnaanacng tcaagaanga caactcctcn 720

cgnttnggca agttcatncg catcaacttn gangtcactg gttacatngt nggngccaan 780

attganacnt atctnctgga aaagtcnvgn gcnatncgnc angcnvgnga ngagagnacn 840

ttncacatct tntactacct gntngcnggn gccaaggana agatganaan tgacntgctn 900

ttggagngct tcaacancta cacattnntn tccaatggct ttgtgcccat cccagcngcn 960

cangatgang agatgttcca gganacnntg gangccatgt cnatcatggg cttcantgaa 1020

gaggancagc tnncnntntt gaaggtngtn tcntcngtcc tncagcttgg aaacatngtc 1080

ttcaagaagg anvgaaacac agaccaggcn tccatgccng anaacacagc ngcncagaaa 1140

gtttgccacc tcntgggnat taangtgaca gatttcacna ganccatcct gaccccncgt 1200

atcaaagttg gacgggangt ngtgcagaan gctcagacna aagaacaggc ngacttcgcn 1260

ntcgaggcnt tngcnaaggc cacntatgan cgccttttcc gntggatnct cagccgtgtn 1320

aacaangccn tggacaagac ccatcggcag ggggcntcct tcctgggnat nctgganatn 1380

gctggntttg anatctttga ggtnaactcc ttcgagcagc tgtgcatcaa ctacaccaac 1440

gagaagctgc agcagctgtt caaccacacn atgttcatcc tggagcagga ngagtaccag 1500

cgngagggca tcgagtggaa cttcatcgac ttcggnctng acctgcagcc nwgyattgag 1560

ctnattgagc ggccgaacaa ccctccnggt gtgctggccc tgctggatga ngantgctgg 1620

ttccccaaag cnacaganaa gtcttttgtg gagaagctnt gcncagagca nggnaancac 1680

cccaanttnc agaagcccaa gcagctnaag gacaaaacng agttctccat catccantan 1740

gctgggaagg tggactacaa ngcnagtgcc tggctgacca agaacatgga cccnctnaat 1800

gacaangtga cntcnctcct caangcctcc tcngacaagt tngtggcnga cctntggaag 1860

gangtggacc gcatngtggg gctggaccag atggccaaga tgacngagag ctcactgccc 1920

agngcctcna agaccaanaa gggcatgttc cgcacngtgg gncagctnta caangagcag 1980

ntggggaanc tgatgncnac nctgcgcaan accacgncna acttngtgcg ctgcatcatc 2040

cccaaccang agaagnggtc nggcaagctg gangcnttnc tngtgctgga ncagctgcgn 2100

tgcaacggng tgntggaagg catccgnatc tgccgncagg gcttccccaa caggatngtc 2160

ttccangagt tccgncaacg ctangagatc ctggcagcna acgccatccc caanggcttc 2220

atggatggna agcaagcctg cattctcatg atcaaagcnc tngaactnga cccnaacntg 2280

tacaggatng ggcagagcaa aatcttcttc cgnacgggng tnctggccca cctngaggag 2340

gancgngacn tgaanatnac ngangtcatc atggccttcc aggcnatgtg tcgtggctac 2400

ctngccvgna aggccttcnc caagnggcag cancagctga cngccatgaa ggtgatccag 2460

aggaactgcg cngcctacct naagctncgn aactggcant ggtggcgnct cttcaccaan 2520

gtnaagccnn tgctncaggt gacacggcag gaggaggaga tgcaggccaa ggaggangag 2580

ntgcanaaga tcanggagcg ncagcagaag gcngaganng agntnvagga gctgvagcag 2640

aagcacacnc agctgncnga ggagaagann ctgctgcagg agcagntgca ggcngagacn 2700

gagctgtang cngagncnga ggagatgcgn gtccggntgg cngccaagaa gcagganctg 2760

gagganatcc tncatgagat ggaggcccgc ctggaggang aggaagaccg gngccagcan 2820

ctncaggcng agaggaagaa gatggcncag cagatgctng acctgganga gcaactggag 2880

gaggangang cngccagnca gaanctacag ctnganaagg tcacngcnga ggccaagatc 2940

aagaanntgg aggangacat cntggtnatg gangatcaga acannaanct ntcaaaagan 3000

cgaaaactcc tngaagagag gntnagngan ttnacaacna anctngcnga ngaggaagan 3060

aaggcnaana acctnacnaa gctgaaganc aagcatgant cnatgatctc aganctggan 3120

gtgnggctga agaangagga gaagagccgg caggagctgg agaanctnaa gnggaanntg 3180

ganggngang ccagtgacnt ccangagcag atcgcngacn tncaggcnca gatngcagag 3240

ctcaagatgc agctggcnaa gaangangan gagctncagg cngcnctngc caggctngan 3300

ganganannn cncagaanaa caangccctn aagaagatnc gngagctnga gggncanatc 3360

tcngacctnc angaggacct ngactcagag cgggcngcca ggaacaaggc nganaagcag 3420

aagcgagacc tnggngagga gctggaggcn ctnaagacng agctgganga nacnctggac 3480

ancacngcna cncagcagga gctcvgngcc aagnggganc aggaggtgac ngtgctgaag 3540

aaggccctgg angangagac ncggtcccat gaggcncagg tccaggagat gaggcagaan 3600

cacncacagg nngtggagga nctcacngag cagctnganc agttcaanag ggccaaggcn 3660

aacctngaca anancaagca gacnctggag aangagaacg cngacctggc nggngagctg 3720

cgngtcctgg gccaggcnaa gcaggaggtg gancanaaga agaagaagct ggaggngcag 3780

ntgcagganc tgcagtccaa gtgcagngat ggggagcgng cccgggcnga gctcanngac 3840

aangtccaca agctncagaa tgaagtngag agngtcacng gnatgctnan ngaggcngag 3900

ggnaangcca tnaanctggc caangangtg gcntccctng gntcccagct ncagganacc 3960

cangagctgc tncaagaaga aacccggcag aagctcaang tgtcnacnaa gctgcgncag 4020

ntggangang annggaacag cctgcangan cagctggang aggagatgga ggcnaagcan 4080

aacctggagc gccanntctc nacnctnaac atncagctct cngactcnaa gaagaagctg 4140

caggactttg cnagnaccnt ngannnyntg ganganggna agaagaggtt ncagaangan 4200

atngagnncc tcanccagca gtaygangag aangcngcng cntanganaa actgganaan 4260

accaagaaca ggctncagca ggagctggan gacctggtng tnganttgga naaccagcgg 4320

caactngtnt ccaanctgga aaagaagcag angaanttng ancagttgtt agcngaggan 4380

aanaacatct cntccaanta ngcggatgan agngacvgag cngangcnga ngcnagggan 4440

aagganacna aggcnntgtc nctngcncgg gccctngang angccntgga ngccaaagan 4500

ganctngagv gnaccaacaa natgctcaan gcnganatgg aagacctngt cagctccaag 4560

gangangtng gcaagaacgt ncatganctg gagaagtcca agcgngccnt ggagacncag 4620

atggangaga tgaanacnca gctggangag yyrgaggang anntgcangc cacngaggan 4680

gccaanctgc ggntngangt caacatgcag gcnctcaang nccagttnga nvgngatctc 4740

cangcncggg angancagaa ngaggagaag aggaggcanc tncagvgnca gctncangag 4800

›INDUSTRIAL APPLICABILITY · 5 of 5

tangagacng aactggaaga ngancgnaag canvgngcnc tggcngcngc agcnaagaag 4860

aagctggang gggacctnaa agacctngag ctncaggcng actcngccat caangggngg 4920

gaggaagcca tcaagcagct ncnnaaactg caggctcaga tgaaggactt ncanagagan 4980

ctggangatg cccgtgcctc cagngangag atctttgcca cnncnaanga gaangagaag 5040

aaagccaaga gnntggangc agacctcatg cagctncaag agganctngc ngcngcngag 5100

agngctcgca ancangcnga cntngagaag gagganctgg cngaggagct ggcnagnagc 5160

ntgtcnggaa ggaanncnct ncaggangag aagcgccgcc tggaggcnng gatcgcncan 5220

ctngaggagg agctggagga ngancagggc aacatggagg cnatgagnga nvgngtncgc 5280

aangcnacnc ngcaggcnga gcanctnagc aangagctgg ccacagancg cagcacngcn 5340

cagaagaatg agagnncncg gcancagctn gagcgncaga acaagganct nmrnagcaag 5400

ntncangagn tnganggngc ngtcaanncc aagntcaant ccacnntngc ggcgctggag 5460

gccaagattg cncagctnga ggagcaggtn gancaggagg ccagagagaa ncaggcggcc 5520

nccaagncgc tgaagcanan ngacaagaag ctnaaggann tnntgctgca ggtggangan 5580

gagcgcaaga tggcngagca gtacaaggag caggcagaga aaggnaannc canggtcaag 5640

cagctnaaga ggcagctgga ngaggcngag gaggagtcnc agngcatcaa cgccaaccgc 5700

aggaagctgc agcgggagct ngangaggcc acngagagca angaggccat gggccgngag 5760

gtgaacgcnc tcaagagcaa nctcaggvga ggaaacgagn cntcnttngt tcctncnaga 5820

aggnctggng gncgtagagt tattgaaaan ncagatggnt ctganganga anbggacnct 5880

cgngacncag acttcaatgg aaccaanncc agtgaatra 5919

1972

›PRT · 1 of 2

Mus musculus

Met Ala Gln Lys Gly Gln Leu Ser Asp Asp Glu Lys Phe Leu Phe Val

1 5 10 15

Asp Lys Asn Phe Met Asn Ser Pro Met Ala Gln Ala Asp Trp Val Ala

20 25 30

Lys Lys Leu Val Trp Val Pro Ser Glu Lys Gln Gly Phe Glu Ala Ala

35 40 45

Ser Ile Lys Glu Glu Lys Gly Asp Glu Val Val Val Glu Leu Val Glu

50 55 60

Asn Gly Lys Lys Val Thr Val Gly Lys Asp Asp Ile Gln Lys Met Asn

65 70 75 80

Pro Pro Lys Phe Ser Lys Val Glu Asp Met Ala Glu Leu Thr Cys Leu

85 90 95

Asn Glu Ala Ser Val Leu His Asn Leu Arg Glu Arg Tyr Phe Ser Gly

100 105 110

Leu Ile Tyr Thr Tyr Ser Gly Leu Phe Cys Val Val Val Asn Pro Tyr

115 120 125

Lys Tyr Leu Pro Ile Tyr Ser Glu Lys Ile Val Asp Met Tyr Lys Gly

130 135 140

Lys Lys Arg His Glu Met Pro Pro His Ile Tyr Ala Ile Ala Asp Thr

145 150 155 160

Ala Tyr Arg Ser Met Leu Gln Asp Arg Glu Asp Gln Ser Ile Leu Cys

165 170 175

Thr Gly Glu Ser Gly Ala Gly Lys Thr Glu Asn Thr Gln Lys Val Ile

180 185 190

Gln Tyr Leu Ala Val Val Ala Ser Ser His Lys Gly Lys Lys Asp Ser

195 200 205

Ser Ile Thr Gly Glu Leu Glu Lys Gln Leu Leu Gln Ala Asn Pro Ile

210 215 220

Leu Glu Ala Phe Gly Asn Ala Lys Thr Val Lys Asn Asp Asn Ser Ser

225 230 235 240

Arg Phe Gly Lys Phe Ile Arg Ile Asn Phe Asp Val Thr Gly Tyr Ile

245 250 255

Val Gly Ala Asn Ile Glu Thr Tyr Leu Leu Glu Lys Ser Arg Ala Ile

260 265 270

Arg Gln Ala Arg Asp Glu Arg Thr Phe His Ile Phe Tyr Tyr Leu Leu

275 280 285

Ala Gly Ala Lys Glu Lys Met Lys Ser Asp Leu Leu Leu Glu Ser Phe

290 295 300

Asn Ser Tyr Thr Phe Leu Ser Asn Gly Phe Val Pro Ile Pro Ala Ala

305 310 315 320

Gln Asp Asp Glu Met Phe Gln Glu Thr Leu Glu Ala Met Ser Ile Met

325 330 335

Gly Phe Asn Glu Glu Glu Gln Leu Ala Ile Leu Lys Val Val Ser Ser

340 345 350

Val Leu Gln Leu Gly Asn Ile Val Phe Lys Lys Glu Arg Asn Thr Asp

355 360 365

Gln Ala Ser Met Pro Asp Asn Thr Ala Ala Gln Lys Val Cys His Leu

370 375 380

Val Gly Ile Asn Val Thr Asp Phe Thr Arg Ala Ile Leu Thr Pro Arg

385 390 395 400

Ile Lys Val Gly Arg Asp Val Val Gln Lys Ala Gln Thr Lys Glu Gln

405 410 415

Ala Asp Phe Ala Ile Glu Ala Leu Ala Lys Ala Thr Tyr Glu Arg Leu

420 425 430

Phe Arg Trp Ile Leu Ser Arg Val Asn Lys Ala Leu Asp Lys Thr His

435 440 445

Arg Gln Gly Ala Ser Phe Leu Gly Ile Leu Asp Ile Ala Gly Phe Glu

450 455 460

Ile Phe Glu Val Asn Ser Phe Glu Gln Leu Cys Ile Asn Tyr Thr Asn

465 470 475 480

Glu Lys Leu Gln Gln Leu Phe Asn His Thr Met Phe Ile Leu Glu Gln

485 490 495

Glu Glu Tyr Gln Arg Glu Gly Ile Glu Trp Asn Phe Ile Asp Phe Gly

500 505 510

Leu Asp Leu Gln Pro Ser Ile Glu Leu Ile Glu Arg Pro Asn Asn Pro

515 520 525

Pro Gly Val Leu Ala Leu Leu Asp Glu Glu Cys Trp Phe Pro Lys Ala

530 535 540

Thr Asp Lys Ser Phe Val Glu Lys Leu Cys Ser Glu Gln Gly Asn His

545 550 555 560

Pro Lys Phe Gln Lys Pro Lys Gln Leu Lys Asp Lys Thr Glu Phe Ser

565 570 575

Ile Ile His Tyr Ala Gly Lys Val Asp Tyr Asn Ala Ser Ala Trp Leu

580 585 590

Thr Lys Asn Met Asp Pro Leu Asn Asp Asn Val Thr Ser Leu Leu Asn

595 600 605

Ala Ser Ser Asp Lys Phe Val Ala Asp Leu Trp Lys Asp Val Asp Arg

610 615 620

Ile Val Gly Leu Asp Gln Met Ala Lys Met Thr Glu Ser Ser Leu Pro

625 630 635 640

Ser Ala Ser Lys Thr Lys Lys Gly Met Phe Arg Thr Val Gly Gln Leu

645 650 655

Tyr Lys Glu Gln Leu Gly Lys Leu Met Ala Thr Leu Arg Asn Thr Thr

660 665 670

Ala Asn Phe Val Arg Cys Ile Ile Pro Asn His Glu Lys Arg Ser Gly

675 680 685

Lys Leu Asp Ala Phe Leu Val Leu Glu Gln Leu Arg Cys Asn Gly Val

690 695 700

Leu Glu Gly Ile Arg Ile Cys Arg Gln Gly Phe Pro Asn Arg Ile Val

705 710 715 720

Phe Gln Glu Phe Arg Gln Arg Tyr Glu Ile Leu Ala Ala Asn Ala Ile

725 730 735

Pro Lys Gly Phe Met Asp Gly Lys Gln Ala Cys Ile Leu Met Ile Lys

740 745 750

Ala Leu Glu Leu Asp Pro Asn Leu Tyr Arg Ile Gly Gln Ser Lys Ile

755 760 765

Phe Phe Arg Thr Gly Val Leu Ala His Leu Glu Glu Glu Arg Asp Leu

770 775 780

Lys Ile Thr Asp Val Ile Met Ala Phe Gln Ala Met Cys Arg Gly Tyr

785 790 795 800

Leu Ala Arg Lys Ala Phe Thr Lys Arg Gln Gln Gln Leu Thr Ala Met

805 810 815

Lys Val Ile Gln Arg Asn Cys Ala Ala Tyr Leu Lys Leu Arg Asn Trp

820 825 830

Gln Trp Trp Arg Leu Phe Thr Lys Val Lys Pro Leu Leu Gln Val Thr

835 840 845

Arg Gln Glu Glu Glu Met Gln Ala Lys Glu Glu Glu Met Gln Lys Ile

850 855 860

Thr Glu Arg Gln Gln Lys Ala Glu Thr Glu Leu Lys Glu Leu Glu Gln

865 870 875 880

Lys His Thr Gln Leu Ala Glu Glu Lys Thr Leu Leu Gln Glu Gln Leu

885 890 895

Gln Ala Glu Thr Glu Leu Tyr Ala Glu Ser Glu Glu Met Arg Val Arg

900 905 910

Leu Ala Ala Lys Lys Gln Glu Leu Glu Glu Ile Leu His Glu Met Glu

915 920 925

Ala Arg Leu Glu Glu Glu Glu Asp Arg Arg Gln Gln Leu Gln Ala Glu

930 935 940

Arg Lys Lys Met Ala Gln Gln Met Leu Asp Leu Glu Glu Gln Leu Glu

945 950 955 960

Glu Glu Glu Ala Ala Arg Gln Lys Leu Gln Leu Glu Lys Val Thr Ala

965 970 975

Glu Ala Lys Ile Lys Lys Leu Glu Asp Asp Ile Leu Val Met Asp Asp

980 985 990

Gln Asn Ser Lys Leu Ser Lys Glu Arg Lys Leu Leu Glu Glu Arg Val

995 1000 1005

Ser Asp Leu Thr Thr Asn Leu Ala Glu Glu Glu Glu Lys Ala Lys Asn

1010 1015 1020

Leu Thr Lys Leu Lys Ser Lys His Glu Ser Met Ile Ser Glu Leu Glu

1025 1030 1035 1040

Val Arg Leu Lys Lys Glu Glu Lys Ser Arg Gln Glu Leu Glu Lys Leu

1045 1050 1055

Lys Arg Lys Leu Glu Gly Asp Ala Ser Asp Phe His Glu Gln Ile Ala

1060 1065 1070

Asp Leu Gln Ala Gln Ile Ala Glu Leu Lys Met Gln Leu Ala Lys Lys

1075 1080 1085

Glu Glu Glu Leu Gln Ala Ala Leu Ala Arg Leu Asp Glu Glu Ile Ala

1090 1095 1100

Gln Lys Asn Asn Ala Leu Lys Lys Ile Arg Glu Leu Glu Gly His Ile

1105 1110 1115 1120

Ser Asp Leu Gln Glu Asp Leu Asp Ser Glu Arg Ala Ala Arg Asn Lys

1125 1130 1135

Ala Glu Lys Gln Lys Arg Asp Leu Gly Glu Glu Leu Glu Ala Leu Lys

1140 1145 1150

Thr Glu Leu Glu Asp Thr Leu Asp Ser Thr Ala Thr Gln Gln Glu Leu

1155 1160 1165

Arg Ala Lys Arg Glu Gln Glu Val Thr Val Leu Lys Lys Ala Leu Asp

1170 1175 1180

Glu Glu Thr Arg Ser His Glu Ala Gln Val Gln Glu Met Arg Gln Lys

1185 1190 1195 1200

His Thr Gln Ala Val Glu Glu Leu Thr Glu Gln Leu Glu Gln Phe Lys

1205 1210 1215

Arg Ala Lys Ala Asn Leu Asp Lys Ser Lys Gln Thr Leu Glu Lys Glu

1220 1225 1230

Asn Ala Asp Leu Ala Gly Glu Leu Arg Val Leu Gly Gln Ala Lys Gln

1235 1240 1245

Glu Val Glu His Lys Lys Lys Lys Leu Glu Val Gln Leu Gln Asp Leu

1250 1255 1260

Gln Ser Lys Cys Ser Asp Gly Glu Arg Ala Arg Ala Glu Leu Ser Asp

›PRT · 2 of 2

1265 1270 1275 1280

Lys Val His Lys Leu Gln Asn Glu Val Glu Ser Val Thr Gly Met Leu

1285 1290 1295

Asn Glu Ala Glu Gly Lys Ala Ile Lys Leu Ala Lys Asp Val Ala Ser

1300 1305 1310

Leu Gly Ser Gln Leu Gln Asp Thr Gln Glu Leu Leu Gln Glu Glu Thr

1315 1320 1325

Arg Gln Lys Leu Asn Val Ser Thr Lys Leu Arg Gln Leu Glu Asp Glu

1330 1335 1340

Arg Asn Ser Leu Gln Asp Gln Leu Asp Glu Glu Met Glu Ala Lys Gln

1345 1350 1355 1360

Asn Leu Glu Arg His Val Ser Thr Leu Asn Ile Gln Leu Ser Asp Ser

1365 1370 1375

Lys Lys Lys Leu Gln Asp Phe Ala Ser Thr Ile Glu Val Met Glu Glu

1380 1385 1390

Gly Lys Lys Arg Leu Gln Lys Glu Met Glu Gly Leu Ser Gln Gln Tyr

1395 1400 1405

Glu Glu Lys Ala Ala Ala Tyr Asp Lys Leu Glu Lys Thr Lys Asn Arg

1410 1415 1420

Leu Gln Gln Glu Leu Asp Asp Leu Val Val Asp Leu Asp Asn Gln Arg

1425 1430 1435 1440

Gln Leu Val Ser Asn Leu Glu Lys Lys Gln Lys Lys Phe Asp Gln Leu

1445 1450 1455

Leu Ala Glu Glu Lys Asn Ile Ser Ser Lys Tyr Ala Asp Glu Arg Asp

1460 1465 1470

Arg Ala Glu Ala Glu Ala Arg Glu Lys Glu Thr Lys Ala Leu Ser Leu

1475 1480 1485

Ala Arg Ala Leu Glu Glu Ala Leu Glu Ala Lys Glu Glu Leu Glu Arg

1490 1495 1500

Thr Asn Lys Met Leu Lys Ala Glu Met Glu Asp Leu Val Ser Ser Lys

1505 1510 1515 1520

Asp Asp Val Gly Lys Asn Val His Glu Leu Glu Lys Ser Lys Arg Ala

1525 1530 1535

Leu Glu Thr Gln Met Glu Glu Met Lys Thr Gln Leu Glu Glu Ser Glu

1540 1545 1550

Asp Asp Val Gln Ala Thr Glu Asp Ala Lys Leu Arg Leu Glu Val Asn

1555 1560 1565

Met Gln Ala Leu Lys Gly Gln Phe Glu Arg Asp Leu Gln Ala Arg Asp

1570 1575 1580

Glu Gln Asn Glu Glu Lys Arg Arg Gln Leu Gln Arg Gln Leu His Glu

1585 1590 1595 1600

Tyr Glu Thr Glu Leu Glu Asp Glu Arg Lys Gln Arg Ala Leu Ala Ala

1605 1610 1615

Ala Ala Lys Lys Lys Leu Glu Gly Asp Leu Lys Asp Leu Glu Leu Gln

1620 1625 1630

Ala Asp Ser Ala Ile Lys Gly Arg Glu Glu Ala Ile Lys Gln Leu Arg

1635 1640 1645

Lys Leu Gln Ala Gln Met Lys Asp Phe Gln Arg Glu Leu Asp Asp Ala

1650 1655 1660

Arg Ala Ser Arg Asp Glu Ile Phe Ala Thr Ser Lys Glu Asn Glu Lys

1665 1670 1675 1680

Lys Ala Lys Ser Leu Glu Ala Asp Leu Met Gln Leu Gln Glu Asp Leu

1685 1690 1695

Ala Ala Ala Glu Arg Ala Arg Lys Gln Ala Asp Leu Glu Lys Glu Glu

1700 1705 1710

Leu Ala Glu Glu Leu Ala Ser Ser Leu Ser Gly Arg Asn Thr Leu Gln

1715 1720 1725

Asp Glu Lys Arg Arg Leu Glu Ala Arg Ile Ala Gln Leu Glu Glu Glu

1730 1735 1740

Leu Glu Glu Glu Gln Gly Asn Met Glu Ala Met Ser Asp Arg Val Arg

1745 1750 1755 1760

Lys Ala Thr Leu Gln Ala Glu Gln Leu Ser Asn Glu Leu Ala Thr Glu

1765 1770 1775

Arg Ser Thr Ala Gln Lys Asn Glu Ser Ala Arg Gln Gln Leu Glu Arg

1780 1785 1790

Gln Asn Lys Glu Leu Arg Ser Lys Leu Gln Glu Val Glu Gly Ala Val

1795 1800 1805

Lys Ala Lys Leu Lys Ser Thr Val Ala Ala Leu Glu Ala Lys Ile Ala

1810 1815 1820

Gln Leu Glu Glu Gln Val Glu Gln Glu Ala Arg Glu Lys Gln Ala Ala

1825 1830 1835 1840

Thr Lys Ser Leu Lys Gln Lys Asp Lys Lys Leu Lys Glu Val Leu Leu

1845 1850 1855

Gln Val Glu Asp Glu Arg Lys Met Ala Glu Gln Tyr Lys Glu Gln Ala

1860 1865 1870

Glu Lys Gly Asn Thr Lys Val Lys Gln Leu Lys Arg Gln Leu Glu Glu

1875 1880 1885

Ala Glu Glu Glu Ser Gln Cys Ile Asn Ala Asn Arg Arg Lys Leu Gln

1890 1895 1900

Arg Glu Leu Asp Glu Ala Thr Glu Ser Asn Glu Ala Met Gly Arg Glu

1905 1910 1915 1920

Val Asn Ala Leu Lys Ser Lys Leu Arg Arg Gly Asn Glu Ala Ser Phe

1925 1930 1935

Val Pro Ser Arg Arg Ala Gly Gly Arg Arg Val Ile Glu Asn Thr Asp

1940 1945 1950

Gly Ser Glu Glu Glu Met Asp Ala Arg Asp Ser Asp Phe Asn Gly Thr

1955 1960 1965

Lys Ala Ser Glu

1970

1972

›PRT · 1 of 2

Oryctolagus cuniculus

Met Ala Gln Lys Gly Gln Leu Ser Asp Asp Glu Lys Phe Leu Phe Val

1 5 10 15

Asp Lys Asn Phe Ile Asn Ser Pro Val Ala Gln Ala Asp Trp Val Ala

20 25 30

Lys Arg Leu Val Trp Val Pro Ser Glu Lys Gln Gly Phe Glu Ala Ala

35 40 45

Ser Ile Lys Glu Glu Lys Gly Asp Glu Val Val Val Glu Leu Val Glu

50 55 60

Asn Gly Lys Lys Val Thr Val Gly Lys Asp Asp Ile Gln Lys Met Asn

65 70 75 80

Pro Pro Lys Phe Ser Lys Val Glu Asp Met Ala Glu Leu Thr Cys Leu

85 90 95

Asn Glu Ala Ser Val Leu His Asn Leu Arg Glu Arg Tyr Phe Ser Gly

100 105 110

Leu Ile Tyr Thr Tyr Ser Gly Leu Phe Cys Val Val Val Asn Pro Tyr

115 120 125

Lys Gln Leu Pro Ile Tyr Ser Glu Lys Ile Val Asp Met Tyr Lys Gly

130 135 140

Lys Lys Arg His Glu Met Pro Pro His Ile Tyr Ala Ile Ala Asp Thr

145 150 155 160

Ala Tyr Arg Ser Met Leu Gln Asp Arg Glu Asp Gln Ser Ile Leu Cys

165 170 175

Thr Gly Glu Ser Gly Ala Gly Lys Thr Glu Asn Thr Lys Lys Val Ile

180 185 190

Gln Tyr Leu Ala Val Val Ala Ser Ser His Lys Gly Lys Lys Asp Thr

195 200 205

Ser Ile Thr Gly Glu Leu Glu Lys Gln Leu Leu Gln Ala Asn Pro Ile

210 215 220

Leu Glu Ala Phe Gly Asn Ala Lys Thr Val Lys Asn Asp Asn Ser Ser

225 230 235 240

Arg Phe Gly Lys Phe Ile Arg Ile Asn Phe Asp Val Thr Gly Tyr Ile

245 250 255

Val Gly Ala Asn Ile Glu Thr Tyr Leu Leu Glu Lys Ser Arg Ala Ile

260 265 270

Arg Gln Ala Arg Glu Glu Arg Thr Phe His Ile Phe Tyr Tyr Leu Ile

275 280 285

Ala Gly Ala Lys Glu Lys Met Arg Asn Asp Leu Leu Leu Glu Gly Phe

290 295 300

Asn Asn Tyr Thr Phe Leu Ser Asn Gly Phe Val Pro Ile Pro Ala Ala

305 310 315 320

Gln Asp Asp Glu Met Phe Gln Glu Thr Val Glu Ala Met Ser Ile Met

325 330 335

Gly Phe Ser Glu Glu Glu Gln Leu Ser Val Leu Lys Val Val Ser Ser

340 345 350

Val Leu Gln Leu Gly Asn Ile Val Phe Lys Lys Glu Arg Asn Thr Asp

355 360 365

Gln Ala Ser Met Pro Asp Asn Thr Ala Ala Gln Lys Val Cys His Leu

370 375 380

Met Gly Ile Asn Val Thr Asp Phe Thr Arg Ser Ile Leu Thr Pro Arg

385 390 395 400

Ile Lys Val Gly Arg Asp Val Val Gln Lys Ala Gln Thr Lys Glu Gln

405 410 415

Ala Asp Phe Ala Val Glu Ala Leu Ala Lys Ala Thr Tyr Glu Arg Leu

420 425 430

Phe Arg Trp Ile Leu Ser Arg Val Asn Lys Ala Leu Asp Lys Thr His

435 440 445

Arg Gln Gly Ala Ser Phe Leu Gly Ile Leu Asp Ile Ala Gly Phe Glu

450 455 460

Ile Phe Glu Val Asn Ser Phe Glu Gln Leu Cys Ile Asn Tyr Thr Asn

465 470 475 480

Glu Lys Leu Gln Gln Leu Phe Asn His Thr Met Phe Ile Leu Glu Gln

485 490 495

Glu Glu Tyr Gln Arg Glu Gly Ile Glu Trp Asn Phe Ile Asp Phe Gly

500 505 510

Leu Asp Leu Gln Pro Cys Ile Glu Leu Ile Glu Arg Pro Asn Asn Pro

515 520 525

Pro Gly Val Leu Ala Leu Leu Asp Glu Glu Cys Trp Phe Pro Lys Ala

530 535 540

Thr Asp Lys Ser Phe Val Glu Lys Leu Cys Thr Glu Gln Gly Asn His

545 550 555 560

Pro Lys Phe Gln Lys Pro Lys Gln Leu Lys Asp Lys Thr Glu Phe Ser

565 570 575

Ile Ile His Tyr Ala Gly Lys Val Asp Tyr Asn Ala Ser Ala Trp Leu

580 585 590

Thr Lys Asn Met Asp Pro Leu Asn Asp Asn Val Thr Ser Leu Leu Asn

595 600 605

Ala Ser Ser Asp Lys Phe Val Ala Asp Leu Trp Lys Asp Val Asp Arg

610 615 620

Ile Val Gly Leu Asp Gln Met Ala Lys Met Thr Glu Ser Ser Leu Pro

625 630 635 640

Ser Ala Ser Lys Thr Lys Lys Gly Met Phe Arg Thr Val Gly Gln Leu

645 650 655

Tyr Lys Glu Gln Leu Gly Lys Leu Met Thr Thr Leu Arg Asn Thr Thr

660 665 670

Pro Asn Phe Val Arg Cys Ile Ile Pro Asn His Glu Lys Arg Ser Gly

675 680 685

Lys Leu Asp Ala Phe Leu Val Leu Glu Gln Leu Arg Cys Asn Gly Val

690 695 700

Leu Glu Gly Ile Arg Ile Cys Arg Gln Gly Phe Pro Asn Arg Ile Val

705 710 715 720

Phe Gln Glu Phe Arg Gln Arg Tyr Glu Ile Leu Ala Ala Asn Ala Ile

725 730 735

Pro Lys Gly Phe Met Asp Gly Lys Gln Ala Cys Ile Leu Met Ile Lys

740 745 750

Ala Leu Glu Leu Asp Pro Asn Leu Tyr Arg Ile Gly Gln Ser Lys Ile

755 760 765

Phe Phe Arg Thr Gly Val Leu Ala His Leu Glu Glu Glu Arg Asp Leu

770 775 780

Lys Ile Thr Asp Val Ile Met Ala Phe Gln Ala Met Cys Arg Gly Tyr

785 790 795 800

Leu Ala Arg Lys Ala Phe Ala Lys Arg Gln Gln Gln Leu Thr Ala Met

805 810 815

Lys Val Ile Gln Arg Asn Cys Ala Ala Tyr Leu Lys Leu Arg Asn Trp

820 825 830

Gln Trp Trp Arg Leu Phe Thr Lys Val Lys Pro Leu Leu Gln Val Thr

835 840 845

Arg Gln Glu Glu Glu Met Gln Ala Lys Glu Asp Glu Leu Gln Lys Ile

850 855 860

Lys Glu Arg Gln Gln Lys Ala Glu Ser Glu Leu Gln Glu Leu Gln Gln

865 870 875 880

Lys His Thr Gln Leu Ser Glu Glu Lys Asn Leu Leu Gln Glu Gln Leu

885 890 895

Gln Ala Glu Thr Glu Leu Tyr Ala Glu Ala Glu Glu Met Arg Val Arg

900 905 910

Leu Ala Ala Lys Lys Gln Glu Leu Glu Glu Ile Leu His Glu Met Glu

915 920 925

Ala Arg Leu Glu Glu Glu Glu Asp Arg Gly Gln Gln Leu Gln Ala Glu

930 935 940

Arg Lys Lys Met Ala Gln Gln Met Leu Asp Leu Glu Glu Gln Leu Glu

945 950 955 960

Glu Glu Glu Ala Ala Arg Gln Lys Leu Gln Leu Glu Lys Val Thr Ala

965 970 975

Glu Ala Lys Ile Lys Lys Leu Glu Asp Asp Ile Leu Val Met Asp Asp

980 985 990

Gln Asn Asn Lys Leu Ser Lys Glu Arg Lys Leu Leu Glu Glu Arg Ile

995 1000 1005

Ser Asp Leu Thr Thr Asn Leu Ala Glu Glu Glu Glu Lys Ala Lys Asn

1010 1015 1020

Leu Thr Lys Leu Lys Asn Lys His Glu Ser Met Ile Ser Glu Leu Glu

1025 1030 1035 1040

Val Arg Leu Lys Lys Glu Glu Lys Ser Arg Gln Glu Leu Glu Lys Leu

1045 1050 1055

Lys Arg Lys Met Asp Gly Glu Ala Ser Asp Leu His Glu Gln Ile Ala

1060 1065 1070

Asp Leu Gln Ala Gln Ile Ala Glu Leu Lys Met Gln Leu Ala Lys Lys

1075 1080 1085

Glu Glu Glu Leu Gln Ala Ala Leu Ala Arg Leu Glu Asp Glu Thr Ser

1090 1095 1100

Gln Lys Asn Asn Ala Leu Lys Lys Ile Arg Glu Leu Glu Gly His Ile

1105 1110 1115 1120

Ser Asp Leu Gln Glu Asp Leu Asp Ser Glu Arg Ala Ala Arg Asn Lys

1125 1130 1135

Ala Glu Lys Gln Lys Arg Asp Leu Gly Glu Glu Leu Glu Ala Leu Lys

1140 1145 1150

Thr Glu Leu Glu Asp Thr Leu Asp Thr Thr Ala Thr Gln Gln Glu Leu

1155 1160 1165

Arg Ala Lys Arg Glu Gln Glu Val Thr Val Leu Lys Lys Ala Leu Asp

1170 1175 1180

Glu Glu Thr Arg Ser His Glu Ala Gln Val Gln Glu Met Arg Gln Lys

1185 1190 1195 1200

His Thr Gln Val Val Glu Glu Leu Thr Glu Gln Leu Glu Gln Phe Lys

1205 1210 1215

Arg Ala Lys Ala Asn Leu Asp Lys Thr Lys Gln Thr Leu Glu Lys Glu

1220 1225 1230

Asn Ala Asp Leu Ala Gly Glu Leu Arg Val Leu Gly Gln Ala Lys Gln

1235 1240 1245

Glu Val Glu His Lys Lys Lys Lys Leu Glu Val Gln Leu Gln Glu Leu

1250 1255 1260

Gln Ser Lys Cys Ser Asp Gly Glu Arg Ala Arg Ala Glu Leu Asn Asp

›PRT · 2 of 2

1265 1270 1275 1280

Lys Val His Lys Leu Gln Asn Glu Val Glu Ser Val Thr Gly Met Leu

1285 1290 1295

Ser Glu Ala Glu Gly Lys Ala Ile Lys Leu Ala Lys Glu Val Ala Ser

1300 1305 1310

Leu Gly Ser Gln Leu Gln Asp Thr Gln Glu Leu Leu Gln Glu Glu Thr

1315 1320 1325

Arg Gln Lys Leu Asn Val Ser Thr Lys Leu Arg Gln Leu Glu Asp Glu

1330 1335 1340

Arg Asn Ser Leu Gln Glu Gln Leu Asp Glu Glu Met Glu Ala Lys Gln

1345 1350 1355 1360

Asn Leu Glu Arg His Ile Ser Thr Leu Asn Ile Gln Leu Ser Asp Ser

1365 1370 1375

Lys Lys Lys Leu Gln Asp Phe Ala Ser Thr Val Glu Ser Leu Glu Glu

1380 1385 1390

Gly Lys Lys Arg Phe Gln Lys Glu Ile Glu Ser Leu Thr Gln Gln Tyr

1395 1400 1405

Glu Glu Lys Ala Ala Ala Tyr Asp Lys Leu Glu Lys Thr Lys Asn Arg

1410 1415 1420

Leu Gln Gln Glu Leu Asp Asp Leu Val Val Asp Leu Asp Asn Gln Arg

1425 1430 1435 1440

Gln Leu Val Ser Asn Leu Glu Lys Lys Gln Lys Lys Phe Asp Gln Leu

1445 1450 1455

Leu Ala Glu Glu Lys Asn Ile Ser Ser Lys Tyr Ala Asp Glu Arg Asp

1460 1465 1470

Arg Ala Glu Ala Glu Ala Arg Glu Lys Glu Thr Lys Ala Leu Ser Leu

1475 1480 1485

Ala Arg Ala Leu Glu Glu Ala Leu Glu Ala Lys Glu Glu Leu Glu Arg

1490 1495 1500

Thr Asn Lys Met Leu Lys Ala Glu Met Glu Asp Leu Val Ser Ser Lys

1505 1510 1515 1520

Asp Asp Val Gly Lys Asn Val His Glu Leu Glu Lys Ser Lys Arg Ala

1525 1530 1535

Leu Glu Thr Gln Met Glu Glu Met Lys Thr Gln Leu Glu Glu Leu Glu

1540 1545 1550

Asp Glu Leu Gln Ala Thr Glu Asp Ala Lys Leu Arg Leu Glu Val Asn

1555 1560 1565

Met Gln Ala Leu Lys Val Gln Phe Glu Arg Asp Leu Gln Ala Arg Asp

1570 1575 1580

Glu Gln Asn Glu Glu Lys Arg Arg Gln Leu Gln Arg Gln Leu His Glu

1585 1590 1595 1600

Tyr Glu Thr Glu Leu Glu Asp Glu Arg Lys Gln Arg Ala Leu Ala Ala

1605 1610 1615

Ala Ala Lys Lys Lys Leu Glu Gly Asp Leu Lys Asp Leu Glu Leu Gln

1620 1625 1630

Ala Asp Ser Ala Ile Lys Gly Arg Glu Glu Ala Ile Lys Gln Leu Leu

1635 1640 1645

Lys Leu Gln Ala Gln Met Lys Asp Phe Gln Arg Glu Leu Glu Asp Ala

1650 1655 1660

Arg Ala Ser Arg Asp Glu Ile Phe Ala Thr Ala Lys Glu Asn Glu Lys

1665 1670 1675 1680

Lys Ala Lys Ser Leu Glu Ala Asp Leu Met Gln Leu Gln Glu Asp Leu

1685 1690 1695

Ala Ala Ala Glu Arg Ala Arg Lys Gln Ala Asp Leu Glu Lys Glu Glu

1700 1705 1710

Leu Ala Glu Glu Leu Ala Ser Ser Leu Ser Gly Arg Asn Ala Leu Gln

1715 1720 1725

Asp Glu Lys Arg Arg Leu Glu Ala Arg Ile Ala Gln Leu Glu Glu Glu

1730 1735 1740

Leu Glu Glu Glu Gln Gly Asn Met Glu Ala Met Ser Asp Arg Val Arg

1745 1750 1755 1760

Lys Ala Thr Gln Gln Ala Glu Gln Leu Ser Asn Glu Leu Ala Thr Glu

1765 1770 1775

Arg Ser Thr Ala Gln Lys Asn Glu Ser Ala Arg Gln Gln Leu Glu Arg

1780 1785 1790

Gln Asn Lys Glu Leu Lys Ser Lys Leu Gln Glu Met Glu Gly Ala Val

1795 1800 1805

Lys Ser Lys Phe Lys Ser Thr Ile Ala Ala Leu Glu Ala Lys Ile Ala

1810 1815 1820

Gln Leu Glu Glu Gln Val Glu Gln Glu Ala Arg Glu Lys Gln Ala Ala

1825 1830 1835 1840

Ala Lys Ala Leu Lys Gln Arg Asp Lys Lys Leu Lys Glu Met Leu Leu

1845 1850 1855

Gln Val Glu Asp Glu Arg Lys Met Ala Glu Gln Tyr Lys Glu Gln Ala

1860 1865 1870

Glu Lys Gly Asn Ala Lys Val Lys Gln Leu Lys Arg Gln Leu Glu Glu

1875 1880 1885

Ala Glu Glu Glu Ser Gln Arg Ile Asn Ala Asn Arg Arg Lys Leu Gln

1890 1895 1900

Arg Glu Leu Asp Glu Ala Thr Glu Ser Asn Glu Ala Met Gly Arg Glu

1905 1910 1915 1920

Val Asn Ala Leu Lys Ser Lys Leu Arg Arg Gly Asn Glu Thr Ser Phe

1925 1930 1935

Val Pro Thr Arg Arg Ser Gly Gly Arg Arg Val Ile Glu Asn Ala Asp

1940 1945 1950

Gly Ser Glu Glu Glu Val Asp Ala Arg Asp Ala Asp Phe Asn Gly Thr

1955 1960 1965

Lys Ser Ser Glu

1970

6644

›DNA · 1 of 2

Oryctolagus cuniculus

agccttggac gccccggcct gggaggtgtg ccagacccgc gctccccgtc cagtttctcc 60

gcgcgccccc cacttggagg ggaccaacca ggcaccatgg cgcagaaggg ccaactcagc 120

gacgatgaga agttcctctt tgtggacaag aacttcatca acagccccgt ggcccaggcc 180

gactgggtgg ccaagaggct ggtgtgggtc ccttcggaga agcagggctt cgaggcggcc 240

agcatcaagg aggagaaggg ggatgaggtg gtcgtggagc tggtggagaa tgggaagaag 300

gtcacggtgg gcaaggatga catccagaag atgaacccgc ccaagttctc caaggtggaa 360

gacatggcgg agctgacgtg tctcaacgaa gcttccgtgc tgcacaacct gagggagagg 420

tacttctctg ggctcatcta cacgtactcc ggcctcttct gcgtggtggt caacccctac 480

aagcagctgc ccatctactc ggagaagatc gtggacatgt acaagggcaa gaagaggcac 540

gagatgccac cacacatcta cgccatcgcc gacacggcct acaggagcat gctgcaggat 600

cgggaggacc agtccattct ctgcacaggc gagtctggag ccgggaagac ggagaacacc 660

aagaaagtca tccagtacct ggccgtggtg gcctcctcgc acaagggcaa gaaggacacg 720

agcatcacgg gggagctgga gaagcagctt ctgcaagcaa accccatcct ggaggccttt 780

ggcaacgcca agacggtcaa gaatgacaac tcctcgcgat tcggcaagtt catccgcatc 840

aactttgacg tcactggtta catcgtgggc gccaacattg agacctatct gctggaaaag 900

tcacgagcca tccgccaagc ccgagaggag aggaccttcc acatctttta ctacctgatt 960

gctggggcca aggagaagat gagaaatgac ttgctcttgg agggcttcaa caactacaca 1020

ttcctctcca atggctttgt gcccatccca gccgcccagg atgacgagat gttccaggaa 1080

acggtggagg ccatgtccat catgggcttc agtgaagagg agcagctgtc tgtgttgaag 1140

gtggtgtctt cagtcctgca gcttggaaac atcgtcttca agaaggaaag aaacacagac 1200

caggcgtcca tgccggacaa cacagctgcc cagaaagttt gccacctcat gggaattaac 1260

gtgacagatt tcaccagatc catcctgacc ccgcgtatca aagttggacg ggacgtagtg 1320

cagaaagctc agacaaaaga acaggcagac ttcgctgtcg aggctttggc aaaggccacg 1380

tatgaacgcc ttttccgctg gatcctcagc cgtgtgaaca aagccctgga caagacccat 1440

cggcaggggg cttccttcct ggggatcctg gacatcgctg gatttgagat ctttgaggtg 1500

aactccttcg agcagctgtg catcaactac accaacgaga agctgcagca gctgttcaac 1560

cacaccatgt tcatcctgga gcaggaggag taccagcgcg agggcatcga gtggaacttc 1620

atcgacttcg ggctcgacct gcagccctgc attgagctca ttgagcggcc gaacaaccct 1680

ccaggtgtgc tggccctgct ggatgaggag tgctggttcc ccaaagccac agataagtct 1740

tttgtggaga agctgtgcac agagcaaggc aaccacccca agttccagaa gcccaagcag 1800

ctcaaggaca aaacggagtt ctccatcatc cattacgctg ggaaggtgga ctacaacgcg 1860

agtgcctggc tgaccaagaa catggacccc ctgaatgaca acgtgacctc cctcctcaac 1920

gcctcctcgg acaagttcgt ggccgaccta tggaaggacg tggaccgcat cgtggggctg 1980

gaccagatgg ccaagatgac agagagctca ctgcccagcg cctccaagac caagaagggc 2040

atgttccgca cggtggggca gctgtacaag gagcagctgg ggaagctgat gaccacgctg 2100

cgcaacacca cgcccaactt cgtgcgctgc atcatcccca accacgagaa gcggtccggc 2160

aagctggacg cgttcctggt gctggagcag ctgcggtgca acggggtgct ggaaggcatc 2220

cgaatctgcc gccagggctt ccccaacagg atcgtcttcc aggagttccg ccaacgctac 2280

gagatcctgg cagccaacgc catccccaag ggcttcatgg atgggaagca agcctgcatt 2340

ctcatgatca aagctctgga actggacccc aacttgtaca ggatcgggca gagcaaaatc 2400

ttcttccgca cgggcgtgct ggcccacctg gaggaggagc gggacttgaa gatcaccgac 2460

gtcatcatgg ccttccaggc catgtgtcgt ggctacctcg cccgcaaggc cttcgccaag 2520

cggcagcagc agctgaccgc catgaaggtg atccagagga actgcgccgc ctacctgaag 2580

ctgcggaact ggcagtggtg gcgcctcttc accaaggtga agccgctgct gcaggtgaca 2640

cggcaggagg aggagatgca ggccaaggag gatgagctgc agaagatcaa ggagcgacag 2700

cagaaggcgg agagcgagct ccaggagctg cagcagaagc acacgcagct gtccgaggag 2760

aagaacctgc tgcaggagca gctgcaggcg gagacggagc tgtacgcgga ggccgaggag 2820

atgcgcgtcc ggctggcggc caagaagcag gagctggagg aaatcctgca tgagatggag 2880

gcccgcctgg aggaggagga agaccggggc cagcagctgc aggccgagag gaagaagatg 2940

gcccagcaga tgctggacct ggaagagcaa ctggaggagg aggaagctgc caggcagaag 3000

ctacagctcg aaaaggtcac cgccgaggcc aagatcaaga agttggagga cgacatcctg 3060

gtcatggacg atcagaacaa caagctctca aaagagcgaa aactcctgga agagaggatt 3120

agtgatttaa caacaaatct tgccgaggag gaagagaagg ccaagaacct gaccaagctg 3180

aagaacaagc atgaatccat gatctcagaa ctggaagtgc ggctgaagaa ggaggagaag 3240

agccggcagg agctggagaa gctgaagcgg aagatggacg gcgaggccag tgacctccac 3300

gagcagatcg ccgacctcca ggcgcagatc gcagagctca agatgcagct ggccaagaag 3360

gaagaggagc tgcaggcggc cctggccagg ctggaggatg aaacgtctca gaagaacaac 3420

gccctgaaga agatccggga gctggagggg cacatctccg acctgcagga ggacctggac 3480

tcagagcggg ccgccaggaa caaggccgag aagcagaagc gagacctggg ggaggagctg 3540

gaggcgctga agacggagct ggaggacacg ctggacacca cggccaccca gcaggagctc 3600

cgggccaagc gggagcagga ggtgacggtg ctgaagaagg ccctggacga ggagacccgg 3660

tcccatgagg cccaggtcca ggagatgagg cagaaacaca cacaggtggt ggaggagctc 3720

acggagcagc tggaacagtt caagagggcc aaggcgaacc tcgacaagac caagcagacg 3780

ctggagaagg agaacgcaga cctggccggc gagctgcggg tcctgggcca ggccaagcag 3840

gaggtggagc acaagaagaa gaagctggag gtgcagctgc aggagctgca gtccaagtgc 3900

agcgatgggg agcgggcccg ggcggagctc aacgacaagg tccacaagct gcagaatgaa 3960

gtggagagcg tcacgggcat gctcagcgag gccgagggga aggccatcaa gctggccaag 4020

gaggtggcgt ccctcgggtc ccagctccag gatacccagg agctgctcca agaagaaacc 4080

cggcagaagc tcaacgtgtc caccaagctg cggcagctgg aggacgagag gaacagcctg 4140

caggagcagc tggacgagga gatggaggcc aagcagaacc tggagcgcca catctccacc 4200

ctgaacatcc agctctccga ctcaaagaag aagctgcagg actttgccag caccgtggag 4260

tccttggagg aaggcaagaa gaggttccag aaggaaattg agagcctcac ccagcagtac 4320

gaagagaaag cagctgctta cgataaactg gaaaagacca agaacaggct tcagcaggag 4380

ctggacgacc tggtcgtaga cttggataac cagcggcaac tggtgtccaa cctggaaaag 4440

aagcagaaga agttcgatca gttgttagcc gaggaaaaga acatctcttc caagtatgcg 4500

gatgaaaggg accgagccga ggctgaagca agggaaaagg aaaccaaggc cttgtccctg 4560

gctcgggccc tcgaggaggc cttggaggcc aaagaggagc tcgagagaac caacaaaatg 4620

ctcaaggccg agatggaaga cctcgtcagc tccaaggacg acgtgggcaa gaacgtccat 4680

gagctggaga agtccaagcg ggccctggag acacagatgg aggagatgaa gacgcagctg 4740

gaagagctag aggacgagct gcaggccacc gaggacgcca agctgcggtt ggaggtcaac 4800

atgcaggccc tcaaagtcca gttcgagcgg gatctccagg cccgggatga gcagaacgag 4860

gagaagagga ggcagctgca gaggcagctg catgagtacg agacggaact ggaagacgag 4920

cgcaagcagc gggccctggc cgcggcagcc aagaagaagc tggaggggga cctgaaagac 4980

ctggagcttc aggcggactc cgccatcaaa gggcgggagg aagccatcaa gcagcttctg 5040

aaactgcagg ctcagatgaa ggacttccag agagaactgg aagatgcccg tgcctccaga 5100

gacgagatct ttgccacagc caaggagaac gagaagaaag ccaagagtct ggaggcagac 5160

›DNA · 2 of 2

ctcatgcagc tacaagagga tctggccgcg gcagagaggg ctcgcaaaca ggcagacttg 5220

gagaaggagg agctggccga ggagctggcc agcagcttgt ccggaaggaa cgcgctgcag 5280

gatgagaagc gccgcctgga ggcccggatc gcacagctgg aggaggagct ggaggaggaa 5340

cagggcaaca tggaggcaat gagcgaccgc gtccgcaagg ctacgcagca ggccgagcag 5400

ctcagcaacg agctggccac agagcgcagc acagcccaga agaatgagag cgcacggcag 5460

cagctcgagc ggcagaacaa ggagctcaag agcaagctgc aggagatgga gggggcagtc 5520

aagtccaagt tcaagtccac tatcgcggcg ctggaggcca agattgcgca gctggaggag 5580

caggttgagc aggaggccag agagaagcag gcggccgcca aggcgctgaa gcagagggac 5640

aagaagctga aggagatgct gctgcaggtg gaagacgagc gcaagatggc tgagcagtac 5700

aaggagcagg cagagaaagg aaacgccaag gtcaagcagc tcaagaggca gctggaggag 5760

gccgaggagg agtcgcagcg catcaacgcc aaccgcagga agctgcagcg ggagctggac 5820

gaggccacgg agagcaacga ggccatgggc cgcgaggtga acgcgctcaa gagcaagctc 5880

aggcgaggaa acgagacctc gttcgttcct accagaaggt ctggagggcg tagagttatt 5940

gaaaacgcag atggctctga ggaggaagtg gacgctcgcg acgcagactt caatggaacc 6000

aaatccagtg aatgaacgac ttcgagtttt gcaccacagc agaaggtccc accaaagaac 6060

agattcaacc aaacccaacc cagcaaaccc tacttagcat cgtccaaccc tgtcttcaag 6120

tttcaaacat cacggacaac cccagaacat aaaaaccact gcctgagtca gggtacagaa 6180

ttccaagttt ttatgatgtg gcagaggaaa aagccaacag gaacaaaaac accctcgctc 6240

taccgatcag gcagagatgt taagtttttt ggaagataca gccacatgaa ccggtcactg 6300

gtcaattcgt ttacatgaca tgggtgaagt tcaccacgat gggtgccgct gccccactcc 6360

cactcccagt tcattcccta acctctgtgc ccttccaagc ccctgagtaa ggccctggtt 6420

tgagaatccc ttctccctct cctggctacc acacctcaga catgcacagt tcaccccgcc 6480

caagtgcctt ctgtagtcac aagaagtaaa aaaggagaca ctgttccatg gatgagaaca 6540

gggacggtcc actgtcttat gtgcacccaa ttgtacttcg gacaccttca ctaataaaag 6600

gtcatacgtc aaaaaaaaaa aaaaaaaaaa ctcgtgccga attc 6644

›Tables in the description — 2

1	6175	DNA	Mus musculus
CDS	(105)...(6020)

2	5919	DNA	Mus musculus
misc_feature	(1)...(5919)	n = A,T,C or G

Claims as granted

16 claims

Classifications

8 codes

IPC · International Patent Classification

Section A — Human necessities

A61K38/00

Section C — Chemistry; metallurgy

C12N1/21
C12N15/12
C07K14/47

USPC · US Patent Classification

514/44435/455435/320.1424/93.2

Claim changes

Soon

Coming soonHow the claims changed between publication and grant

See which claims were amended, added or cancelled during examination, with every added and removed word marked.

AmendedAddedCancelledUnchanged

The published claims of this application are not paired with the granted ones in what we hold.

File wrapper

⤢ drag to zoom

USPTOApplicanthover for detail · click to open

Pendency

7.5 y

2,728 days filing → grant

Office actions

non-final + final

Responses

1 RCE

Interviews

examiner interview summaries

Examiner

Scott D. Priebe

art unit 1632 · TC 1600

Law firm

Law firm · Log in to unlock

Citations: 12 back · 1 forward

See the full prosecution history — every USPTO and applicant action on this file, in order.

Documents

Log in to open the documents of this file: the application as filed, every office action and response, the notice of allowance.

Chain of title

⤢ drag to zoom

Titlehover for detail · click to open

See the full assignment history — every owner this patent has passed through, with recordation dates and reel/frame numbers.