Abstract:
The present invention relates to a novel protein isolated from the leukocyte of IgA nephropathy patients, and DNA encoding the protein. It also relates to an oligonucleotide based on the nucleotide sequence complementary with the DNA, an antibody which specifically reacts with the protein and, furthermore, diagnostic and therapeutic drugs of IgA nephropathy comprising it.

Description:
CROSS-REFERENCE TO RELATED APPLICATION 
     This is a continuation-in-part application of PCT/JP97/04469 filed on Dec. 5, 1997. 
    
    
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The present invention relates to a novel protein whose expression level fluctuates in leukocytes of IgA nephropathy patients in comparison with leukocytes of healthy persons, an antibody against the protein, DNA encoding the protein, methods for detecting the protein using the antibody, mRNA and DNA using a primer comprising a complementary nucleotide sequence to the DNA, and diagnosis and treatment of IgA nephropathy. 
     2. Brief Description of the Background Art 
     IgA nephropathy is a chronic glomerulonephritis which is characterized in that an IgA immune complex considered to be originated from blood deposits in glomerulus of the kidney. In Japan, the IgA nephropathy occupies 30% or more of primary renal diseases, and is the most common renal disease. Moreover, 15 to 30% of the patients with IgA nephropathy achieve renal failure due to poor prognosis. However, since the underlying cause of IgA nephropathy is still unclear, a fundamental therapeutic method has not been found. Additionally, definite diagnosis of IgA nephropathy imposes heavy burden on patients, because the method is carried out by removing a portion of the kidney by biopsy and recognizing deposition of the IgA immune complex in mesangium by means of an immunological staining. 
     It has been reported that about 50% of the patients with IgA nephropathy have a high blood IgA level [ Diseases of the Kidney,  5th edition (1993),  Nephron,  29, 170 (1981)]. It is considered that B cells relate to the production of IgA in blood and T cells relate to the regulation of the production. Furthermore, it has been reported that the production of cytokine, such as interleukin 4, interleukin 5, interleukin 6 or TGF-β (transforming growth factor-β), is high in peripheral T cells of IgA nephropathy patients in comparison with healthy persons [ Clinical  &amp;  Experimental Immunology,  103, 125 (1996),  Kidney International,  46, 862 (1994))] and that integrin, such as VLA (very late activation)-4 and VLA-5, are strongly activated in peripheral lymphocytes of IgA nephropathy patients [ Nephrology, Dialysis, Transplantation,  10, 1342 (1995)]. 
     On the basis of these facts, it is considered that, in IgA nephropathy, excessive IgA is produced due to abnormality in the immune system, the resulting IgA immune complex in blood deposits on the glomerulus, and the complement system is activated on the deposited IgA immune complex and the like to exert influence and cause disorders of the glomerulus. However, the cause of IgA nephropathy has not yet been determined. 
     SUMMARY OF THE INVENTION 
     Elucidation of the cause of IgA nephropathy, as well as a treatment or diagnosis which can reduce a burden on patients are long-sought. The present invention provides a novel protein which has been determined to have increased expression level in leukocytes of IgA nephropathy patients, as well as an antibody against the protein, DNA encoding the protein, methods for detecting the protein using the antibody, mRNA and DNA using the oligonucleotide comprising a complementary nucleotide sequence to the DNA, and diagnostic and therapeutic agents of IgA nephropathy. 
     The present invention relates to a protein comprising the amino acid sequence represented by SEQ ID NO:2; DNA encoding the protein of the present invention; DNA which comprises the nucleotide sequence represented by SEQ ID NO:1; and DNA which hybridizes with the DNA comprising the above nucleotide sequence under stringent conditions. Furthermore, the present invention relates to a recombinant vector which comprises the DNA and a vector; a transformant obtained by introducing the recombinant vector into a host cell; and a process for producing the protein, comprising culturing the transformant in a medium to produce and accumulate the protein of the present invention in the culture, and recovering the protein from the resulting culture. 
     Moreover, the present invention relates to a method for detecting the mRNA derived from the protein of the present invention using an oligonucleotide comprising a portion of the nucleotide sequence of the DNA of the present invention or a portion of the nucleotide sequence complementary to the DNA, for example, the oligonucleotide represented by SEQ ID NO:6 or NO:7; and IgA nephropathy diagnostic agents comprising the oligonucleotide. 
     Also, the present invention relates to a method for inhibiting expression of the protein of the present invention using an oligonucleotide comprising a portion of the nucleotide sequence of the DNA of the present invention or a portion of the nucleotide sequence complementary to the DNA, for example, the oligonucleotide represented by SEQ ID NO:6 or NO:7; and IgA nephropathy therapeutic agents comprising the oligonucleotide. 
     Additionally, the present invention relates to an antibody which specifically reacts with the protein of the present invention; a method for immunologically detecting the protein of the present invention using the antibody of the present invention; and IgA nephropathy diagnostic and therapeutic agents comprising the antibody. 
    
    
     BRIEF EXPLANATION OF THE DRAWINGS 
     FIG. 1 shows an autoradiograph of the Northern blot of KIAA0235 in human tissue and cell in Example 5. Both A and B are results of human tissue, wherein A represents the autoradiograph of the filter blotting each RNA of  1 : pancreas,  2 : kidney,  3 : skeletal muscle,  4 : Liver,  5 : lung,  6 : placenta,  7 : brain, and  8 : heart, and B represents the autoradiograph of the filter blotting each RNA of  1 : peripheral leukocyte,  2 : colon,  3 : small intestine,  4 : ovary,  5 : testis,  6 : prostate,  7 : thymus, and  8 : spleen. C is a result of human cell line, which represents the autoradiograph of the filter blotting the RNA of  1 : HeLa and  2 : KG-1. Lines at the leftmost end of A correspond to the RNA marker positions indicating 9.5 kb, 7.5 kb, 4.4 kb, 2.4 kb and 1.35 kb from the top. 
     FIG. 2 shows the detection results of RT-PCR of KIAA0235 against the leukocyte of IgA nephropathy patients and healthy persons by the fluorimager in Example 4.  1  to  5  show the RT-PCR of the leukocyte of 5 healthy persons,  6  to  10  show that of 5 IgA nephropathy patients, and C shows PCR using as the template the plasmid to which the amplified fragment of the KIAA0235 cDNA obtained by differential display is introduced as the positive control. 
    
    
     DETAILED DESCRIPTION OF THE INVENTION 
     This application is based on Japanese application No. 8-325752 filed on Dec. 5, 1996 and PCT/JP97/04469 filed on Dec. 5, 1997, the entire contents of which are incorporated hereinto by reference. 
     The protein of the present invention includes, for example, a protein comprising the amino acid sequence represented by SEQ ID NO:2. The DNA of the present invention includes a DNA sequence encoding the protein of the present invention, a DNA sequence comprising the nucleotide sequence represented by SEQ ID NO:1, a DNA sequence which hybridizes with the DNA under stringent conditions, and the like. 
     The DNA encoding the protein of the present invention includes DNAs having a nucleotide sequence different from the nucleotide sequence represented by SEQ ID NO:1 because generally there are various genetic codes per one amino acid. Those of ordinary skill in this art are well-aware of how to substitute one or more degenerate codons to create an equivalent DNA encoding the same protein sequence. This includes, for example, selecting degenerate codons which may be more common in a particular expression system, such as yeast. 
     Furthermore, the DNA which hybridizes under stringent conditions with a DNA comprising a nucleotide sequence represented by SEQ ID NO:1 means a DNA in which a mutation, such as substitution, deletion, insertion, addition and the like, is introduced at least one portion within the range that the inherent activities of the protein are not lost, and a DNA which is obtained by colony hybridization or plaque hybridization [ Molecular Cloning, A Laboratory Manual,  Second Edition (edited by Sambrook, Fritsch and Maniatis, Cold Spring Harbor Laboratory Press (1989)) (referred to as “ Molecular Cloning, A Laboratory Manual,  2nd ed.” hereinafter) using, as a probe, a DNA comprising a nucleotide sequence represented by SEQ ID NO:1 or a fragment thereof. 
     The preparation and expression of the DNA encoding the novel protein of the present invention is carried out according to the process described in  Molecular Cloning, A Laboratory Manual,  2nd ed.,  Current Protocols in Molecular Biology, Supplement  1  to  34 (edited by Ausubel, Brent, Kingston, Moore, Seidman, Smith and Struhl, published by Green Publishing Associates and Wiley-Interscience, 1987-1996 edition) (referred to as “ Current Protocols in Molecular Biology, Supplement  1  to  34”), and the like. 
     In the present invention, in order to obtain a novel protein, taking note of the difference in the expression quantity of mRNA in leukocytes between patients with IgA nephropathy and healthy persons, the differential display method [ FEBS Letters,  351, 231 (1994)] is used. The differential display method is a method in which cloning of a novel gene is carried out using pattern of expression as an index. That is, an amplified cDNA fragment of a novel gene whose expression level increases or decreases significantly in leukocytes of a patient with IgA nephropathy as compared with leukocytes of a healthy person is obtained by subjecting total RNA or mRNA extracted from cells to the polymerase chain reaction (PCR) using various primers. At the same time, a cDNA library is constructed from human undifferentiated myeloid cell line KG-1, and all nucleotide sequences of cDNA of respective clone are determined to form a database beforehand. The homology between the nucleotide sequence of the clone therein and the above-described amplified cDNA fragment is identified, and a novel full-length cDNA is obtained. This method is described below. 
     Examples of the method for the preparation of a total RNA from leukocytes of patients with IgA nephropathy and leukocytes of healthy persons include guanidine thiocyanate-cesium trifluoroacetate method [ Methods in Enzymol.,  154, 3 (1987)], the AGPC method ( Jikken Igaku,  9, 1937 (1991)), RNeasy kit for recovering RNA (produced by QIAGEN), and the like. 
     Examples of the method for preparing poly(A) +  RNA from the total RNA include oligo(dT)-immobilized cellulose column method ( Molecular Cloning, A Laboratory Manual,  2nd ed.) and the like. Also, examples of the kit for preparing mRNA from leukocytes of patients with IgA nephropathy and leukocytes of healthy persons include Fast Track mRNA Isolation Kit (manufactured by Invitrogen), Quick Prep mRNA Purification Kit (manufactured by Pharmacia), and the like. 
     Using an anchor primer, cDNA is synthesized in the usual way from the RNA extracted by the above-described method from leukocytes of a patient with IgA nephropathy or leukocytes of a healthy person, and PCR is carried out using an anchor primer having a 5′-end labeled with fluorescence and an arbitrary primer. The anchor primer is a primer in which an oligonucleotide of adenine, guanine or cytosine, excluding thymidine, is added to the 3′-end of an oligo(dT) sequence which hybridizes with a 3′-end poly(A) sequence of mRNA. The arbitrary primer is typically an oligonucleotide which amplifies various cDNA sequences and can yield a large number of amplified cDNA fragments by a single reaction. Preferably, the oligonucleotide has a length of about 10 mer. 
     After the PCR, each of the amplified cDNA is subjected to polyacrylamide gel electrophoresis, and the electrophoresis pattern of the amplified cDNA fragment is compared by detecting the fluorescence with a fluorimager. The cDNA fragment which can be amplified by only leukocytes of a patient with IgA nephropathy is extracted from the gel, the amplified cDNA fragment is inserted into a vector, and the nucleotide sequence of the DNA is determined by a usually used nucleotide sequence analyzing method such as the dideoxy method of Sanger et al.  [Proc. Natl. Acad. Sci. USA,  74, 5463 (1977)], or like. 
     Examples of the vector to which the DNA fragment is inserted include pDIRECT [ Nucleic Acids Research,  18, 6069 (1990)], pPCR-Script Amp SK(+) [manufactured by Stratagene,  Strategies,  5, 6264 (1992)], pT7Blue (manufactured by Novagen), pCR II [manufactured by Invitrogen,  Biotechnology,  9, 657 (1991)], pCR-TRAP (manufactured by Genehunter), pNoTA T7  (manufactured by 5′→3′) and the like. 
     The analysis of the nucleotide sequence is carried out by using a nucleotide sequence automatic analyzer, such as 373A•DNA (manufactured by Applied Biosystems), and the like. 
     Full-length cDNA can be obtained by carrying out screening according to hybridization using the above-described amplified cDNA fragment as the probe and various cDNA libraries. Also, as in the present invention a clone having the full-length cDNA can be obtained by constructing a cDNA library in advance, determining all nucleotide sequences of a respective clone to form a data base, and then screening a clone having the nucleotide sequence of the above-described amplified cDNA fragment. The method for preparing a cDNA library will be described. 
     Examples of the method for the preparation of the cDNA library include methods described in  Molecular Cloning, A Laboratory Manual,  2nd. ed., or  Current Protocols in Molecular Biology, Supplement  1  to  34, methods using a commercially available kit, such as Super Script™ Plasmid System for cDNA Synthesis and Plasmid Cloning (manufactured by Life Technologies) or ZAP-cDNA Synthesis Kit (manufactured by Stratagene), and the like. 
     In the preparation of the cDNA library, the vector to which the cDNA, synthesized using mRNA extracted from human undifferentiated myeloid IgA line KG-1 as a template, is inserted may be any vector so long as the cDNA can be inserted thereto. Examples include ZAP Express [ Strategies,  5, 58 (1992)], pBluescript II SK(+) [ Nucleic Acids Research,  17, 9494 (1989)], λ zap II (manufactured by Stratagene), λgt10, λgt11  [DNA Cloning, A Practical Approach,  1, 49, (1985)], Lambda BlueMid (manufactured by Clonetech), λExCell, pT7T318U (manufactured by Pharmacia), pcD2  [Mol. Cell. Biol.,  3, 280 (1983)], pUC18  [Gene,  33, 103 (1985)], and the like. 
     With regard to the  Escherichia coli  for introducing the cDNA library constituted by the vector, any microorganism belonging to  Escherichia coli  can be used so long as the introduction, expression and maintenance of the cDNA library can be conducted. Examples include  Escherichia coli  KL1-Blue MRF′ [ Strategies,  5, 81, (1992)],  Escherichia coli  C600  Genetics,  39, 440 (1954)],  Escherichia coli  Y1088,  Escherichia coli  Y1090 [ Science,  222, 778 (1983)],  Escherichia coli  NM522 [ J. Mol. Biol.,  166, 1 (1983)],  Escherichia coli  K802 [ J. Mol. Biol.,  16, 118 (1996)],  Escherichia coli  JM105 [ Gene,  38, 275 (1985)], and the like. 
     The cDNA can be also obtained without preparing a cDNA library by the 5′-RACE (rapid amplification of cDNA ends) and 3′-RACE [ Proc. Natl. Acad. Sci. USA,  85, 8998 (1988)] in which adapters are added to both ends of the cDNA and then PCR is carried out using primers based on the nucleotide sequence of the adapter and the nucleotide sequence of the amplified fragment. Alternatively, the cDNA can be obtained by PCR based on the nucleotide sequence represented by SEQ ID NO:1 or a chemical synthesis method using a DNA synthesizer. 
     A cDNA clone can be selected from the cDNA library according to a colony hybridization or plaque hybridization method ( Molecular Cloning, A Laboratory Manual,  2nd ed.) using a probe labeled with an isotope or fluorescence. The cDNA may be also prepared according to the polymerase chain reaction (PCR) [ Molecular Cloning, A Laboratory Manual,  2nd ed. or  Current Protocols in Molecular Biology, Supplement  1  to  34] by preparing a primer and using, as a template, cDNA synthesized from poly (A) + RNA or mRNA, or cDNA library. 
     The nucleotide sequence of the DNA can be determined by cleaving the cDNA clone selected by the above method with an appropriate restriction enzyme, cloning to a plasmid, such as pBluescript KS(+) (manufactured by Stratagene) or the like, and then analyzing by a conventional nucleotide sequence analysis method, such as dideoxy method of Sanger et al. [ Proc. Natl. Acad. Sci., U.S.A.,  74, 5463 (1977)] or the like. The nucleotide sequence can be analyzed by using a nucleotide sequence automatic analyzer, such as 373A•DNA sequencer (manufactured by Applied Biosystems) or the like. 
     A transformant which expresses the protein of the present invention can be obtained by preparing a transformed vector to which the full-length DNA prepared according to the above method is inserted into a downstream site of the promoter in an appropriate vector. 
     As the host cell, any bacterium, yeast, animal cell, insect cell, and the like, can be used so long as they can express the gene of interest. Examples of the bacterium include bacteria belonging to the genus Escherichia, Serratia, Corynebacterium, Brevibacterium, Pseudomonas, Bacillus, Microbacterium and the like, for example  Escherichia coli, Bacillus subtilis, Bacillus amyloliquefaciens, Brevibacterium flavum, Brevibacterium lactofermentum, Corynebacterium glutamicum, Microbacterium ammoniaphilum,  and the like. Examples of the yeast include  Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactis, Trichosporon pullulans, Schwanniomyces alluvius  and the like. Examples of the animal cell include human Namalwa cell, monkey COS cell, Chinese hamster CHO cell, and the like. Examples of the insect cell include  Spodoptera frugiperda  oocytes Sf9 and Sf21 ( Bacurovirus Expression Vectors, A Laboratory Manual,  Oreilly, Miller and Luckow, W.H. Freeman and Company, New York (1992) (referred to as “ Bacurovirus Expression Vectors, A Laboratory Manual”  hereinafter)),  Trichoplusia ni  oocyte Tn5 (High 5, manufactured by Pharmigen), and the like. 
     Any vector can be used as the vector to which the DNA of the present invention is inserted so long as it can introduce the DNA and drive the expression in the host cell. 
     When a bacterium, such as  Escherichia coli,  is used as the host cell, it is preferred that the vector is constituted by a promoter, a ribosome binding sequence, the DNA of the present invention and a transcription termination sequence. A promoter controlling gene may be also contained. 
     Examples of the expression vector include pKYP10 (Japanese Published Unexamined Patent Application No. 110600/83), pLSA1  [Agric. Biol. Chem.,  53, 277 (1989)], pGEL1 [ Proc. Natl. Acad. Sci. USA,  82, 4306 (1985)], and the like. 
     With regard to the promoter, any promoter can be used so long as it can drive the expression in the host cell. Examples include promoters originated from  Escherichia coli,  phage and the like (for example, trp promoter (Ptrp), lac promoter (Plac), T7 lac promoter, PL promoter, PR promoter, and the like). Also, artifically designed and modified promoters, such as a promoter in which two Ptrp are linked in series (Ptrp×2), tac promoter, and the like, can be used. 
     With regard to the ribosome binding sequence, it is preferred to use a plasmid in which the space between Shine-Dalgarno sequence (referred to as “SD sequence” hereinafter) and the initiation codon is adjusted to an appropriate distance (for example, 6 to 18 bases). 
     With regard to the recombinant rector of the present invention, it is preferred to substitute a suitable nucleotide in order that the nucleotide sequence of the DNA of the present invention forms a codon suitable for the expression of a host cell. 
     The transcription termination sequence is not always necessary for the recombinant vector of the present invention. However, it is preferred to arrange the transcription terminating sequence just downstream of the structural gene. 
     With regard to the method for the introduction of the recombinant vector to the bacterium, any one of the known methods for introducing DNA into the bacterium, such as a method in which calcium ion is used [ Proc. Natl. Acad. Sci. USA,  69, 2110 (1972)], a protoplast method (Japanese Published Unexamined Patent Application No. 2483942/88), and the like, can be used. 
     When yeast is used as the host cell, YEp13 (ATCC 37115), YEp24 (ATCC 37051), YCp50 (ATCC 37419), or the like is used as the expression vector. Any promoter can be used so long as it can drive the expression in yeast. Examples include promoters of genes in the glycolysis system (for example, hexosekinase, and the like), gal 1 promoter, gal 10 promoter, heat shock protein promoter, MFα1 promoter, CUP 1 promoter and the like. 
     With regard to the method for the introduction of the recombinant vector, any one of known methods for introducing DNA into yeast, such as an electroporation method [ Methods. Enzymol.,  194, 182-187 (1990)], a spheroplast method [ Proc. Natl. Acad. Sci. USA,  84, 1929-1933 (1978)], a lithium acetate method [ J. Bacteriol.,  153, 163-168 (1983)], and the like can be used. 
     When animal cells are used as the host cells, pAGE107 [Japanese Published Unexamined Patent Application No. 22979/91;  Cytotechnology,  3, 133 (1990)], pAS3-3 (Japanese Published Unexamined Patent Application No. 227075/90), pAMoERC3Sc, pcDM8  [Nature,  329, 840 (1987)], pcDNAI/Amp, pcDNAI (both manufactured by Funakoshi), and the like can be exemplified as the expression vector. Any promoter can be used so long as it can drive the expression in animal cell. Examples include a promoter of IE (immediate early) gene of cytomegalovirus (CMV), a promoter of SV40 or metallothionein, and the like. Also, the enhancer of the IE gene of human CMV may be used together with the promoter. 
     With regard to the method for the introduction of the recombinant vector into animal cells, any one of the known methods for introducing DNA into animal cells, such as an electroporation method [ Cytotechnology,  3, 133 (1990)], a calcium phosphate method (Japanese Published Unexamined Patent Application No. 227075/90), a lipofection method [ Proc. Natl. Acad. Sci. USA,  84, 7413 (1987)], and the like can be used. 
     When an insect cell is used as the host cell, the protein can be expressed by known methods described in, for example,  Current Protocols in Molecular Biology, supplement  1-34; Bacurovirus Expression Vectors, A Laboratory Manual; or the like. That is, a recombinant gene transfer vector and bacurovirus are simultaneously introduced into an insect cell to obtain a recombinant virus in an insect cell culture supernatant, and then insect cells are infected with the thus obtained recombinant virus to obtain protein expression insect cell. 
     Examples of the gene transfer vector include pVL1392, pVL1393, pBlueBacIII (all manufactured by Invitrogen), and the like. 
     Examples of the bacurovirus include Autographa californica nuclear polyhedrosis virus with which insects of the family Barathra are infected, and the like. 
     The method for the co-transfer of the above-described recombinant gene transfer vector and the above-described bacurovirus for the preparation of the recombinant virus include a calcium phosphate method (Japanese Published Unexamined Patent Application No. 227075/90), a lipofection method [ Proc. Natl. Acad. Sci. USA,  84, 7413 (1987)], and the like. 
     With regard to the gene expression method, a secreted protein production, a fusion protein expression and the like can be effected in accordance with the method described in J. Sambrook et al. ( Molecular Cloning,  2nd, ed.) in addition to the direction expression. 
     When expressed in a yeast, an animal cell or a insect cell, a protein to which sugar or sugar chain is added by exo- and endo-glycosidase can be obtained. 
     The protein of the present invention can be produced by culturing the thus obtained transformant in a culture medium to produce and accumulate the protein of the present invention, and recovering the protein from the resulting culture. Culturing of the transformant of the present invention in a culture medium is carried out in accordance with a usual method used in culturing of host cells. 
     The medium for culturing the transformant obtained by using as the host cell a microorganism, such as  Escherichia coli,  yeast or the like, may be either a natural medium or a synthetic medium, so long as it contains a suitable carbon source, a suitable nitrogen source, and a suitable inorganic salt and the like, which enables culturing the transformant efficiently. 
     Examples of the carbon source include carbohydrates (for example, glucose, fructose, sucrose, molasses, starch, starch hydrolysate, and the like), organic acids (for example, acetic acid, propionic acid, and the like), and alcohols (for example ethanol, propanol, and the like). 
     Examples of the nitrogen source include ammonia, various ammonium salts of inorganic acids or organic acids (for example, ammonium chloride, ammonium sulfate, ammonium acetate, ammonium phosphate, and the like), other nitrogen-containing compounds, peptone, meat extract, yeast extract, corn steep liquor, casein hydrolysate, soybean meal and soybean meal hydrolysate, various fermented cells and hydrolysates thereof, and the like. 
     Examples of inorganic substance include potassium dihydrogen phosphate, dipotassium hydrogen phosphate, magnesium phosphate, magnesium sulfate, sodium chloride, ferrous sulfate, manganese sulfate, copper sulfate, calcium carbonate, and the like. 
     The culturing is carried out under aerobic conditions by means of shaking, aeration stirring or the like at 15 to 45° C. for 16 to 96 hours. The pH of the medium is maintained at 3.0 to 9.0 during the culturing. Adjustment of the medium pH is carried out using an inorganic or organic acid, an alkali solution, urea, calcium carbonate, ammonia and the like. 
     Also, antibiotics (for example, ampicillin, tetracycline, and the like) may be added to the medium during the culturing as occasion demands. 
     When a microorganism transformed with an expression vector containing an inducible promoter is cultured, an inducer may be added to the medium as occasion demands. For example, isopropyl-β-D-thiogalactopyranoside (IPTG) or the like may be added to the medium when a microorganism transformed with an expression vector containing lac promoter is cultured, or indoleacrylic acid (IAA) or the like may by added thereto when a microorganism transformed with an expression vector containing trp promoter is cultured. 
     Examples of the medium used in the culturing of a transformant obtained using an animal cell as the host cell include RPMI 1640 medium, Eagle&#39;s MEM medium, and any one of these media further supplemented with fetal calf serum. The culturing is carried out generally at a temperature of 35 to 37° C. for a period of 3 to 7 days in the presence of 5% CO 2 . As occasion demands, antibiotics (for example, kanamycin, penicillin, and the like) may be added to the medium during the culturing. 
     Examples of the medium used in the culturing of a transformant obtained using an insect cell as the host cell include TNM-FH medium (manufactured by Pharmingen), Sf900 II SFM (manufactured by Life Technologies), ExCell 400 or ExCell 405 (both manufactured by JRH Biosciences), and the like. The culturing is carried out generally at a temperature of 25 to 30° C. for a period of 1 to 4 days. Additionally, antibiotics (for example, gentamicin, and the like) may be added to the medium during the culturing as occasion demands. 
     When the protein of the present invention is expressed in a dissolved state inside the cells, the cells after completion of the culturing are recovered by centrifugation, suspended in an aqueous buffer and then disrupted by ultrasonic, French press or the like to obtain the protein from a supernatant fluid prepared by centrifugation. Also, when the protein forms an inclusion body, the inclusion body is solubilized using a protein denaturing agent, and then the solubilized solution is diluted to or dialyzed against a solution containing no protein denaturing agent or a dilute solution containing a protein denaturing agent in such a concentration that the protein is not denatured in order to form a renatured protein. 
     When the protein of the present invention or a derivative thereof, such as a sugar-modified product or the like, is secreted outside the cells, the protein or the derivative, such as a sugar-modified product or the like, can be recovered from the culture supernatant. That is, the isolation and purification can be conducted by using isolation steps, such as solvent extraction, fractional precipitation by an organic solvent, salting-out, dialysis, centrifugation, ultrafiltration, ion exchange chromatography, gel filtration chromatography, hydrophobic interaction chromatography, affinity chromatography, reverse, phase chromatography, crystallization, electrophoresis, and the like, alone or as a combination thereof. 
     Furthermore, the protein of the present invention can be prepared according to a chemical synthesis method based on the amino acid sequence represented by SEQ ID NO:2. 
     Examples of a method for detecting the mRNA of a novel protein using the oligonucleotide based on the nucleotide sequence of the DNA of the present invention includes Northern hybridization [ Molecular Cloning, A Laboratory Manual,  2nd ed., Cold Spring Harbor Laboratory Press (1989)], PCR [ PCR Protocols,  Academic Press (1990)], and the like. Particularly, RT (Reverse Transcribed)-PCR is simple and easy and can therefore be applied to the diagnosis of IgA nephropathy. Specifically, the amplified fragment is detected by collecting blood from human to recover leukocyte, transforming the RNA isolated therefrom into cDNA using an oilgo(dT) primer and a reverse transcriptase into, and conducting PCR using a pair of oligonucleotide primers corresponding to the mRNA to be detected. 
     Examples of the oligonucleotide primers include a sense primer corresponding to the 5′-end side nucleotide sequence, and an antisense primer corresponding to the 3′-end side nucleotide sequence, of a portion of the mRNA to be detected. In this case, the nucleic acid corresponding to uracil in mRNA corresponds to thymidine in the oligonucleotide primer. 
     As the sense primer and antisense primer, it is preferred to use oligonucleotides in which melting point (T s ) and the number of bases are not significantly different from each other. Preferably, the base number is 15 to 40 mer. 
     The nucleotide sequence moiety to be amplified using the above oligonucleotide primer may be any nucleotide sequence region of the mRNA, but a nucleotide sequence region which has a length of 50 bp to 2 kbp and does not contain a sequence rich in a repeating sequence or GC (guanine-cytosine) bases is preferred. 
     Furthermore, the expression of the protein can be inhibited by repressing the transcription of DNA or translation of mRNA using an antisense RNA/DNA [ Chemistry,  46, 681 (1991),  Biotechnology,  9, 358 (1992)]. The inhibition of production of the protein using anti-sense RNA/DNA technology can be carried out by designing and preparing an oligonucleotide based on the nucleotide sequence of a portion of the DNA encoding the protein of the present invention, preferably that of 10 to 50 bases positioned in the translation initiation site, and administrating it in vivo. As the nucleotide sequence of the synthetic oligonucleotide, those which partially comprises the nucleotide sequence of the antisense chain of the DNA encoding of the protein of the present invention, or those which have been modified to the extent not to lose the activity of inhibiting the expression of the protein activity can be used. As oligonucleotide, DNA, RNA or their derivatives, such as methyl or phosphorothioate derivatives, can be used. 
     The antibody can be produced by immunizing an animal using the protein of the present invention as an antigen or a peptide, chemically synthesized based on the amino acid sequence represented by SEQ ID NO:2 which is a protein of the present invention. A monoclonal antibody to the protein of the present invention can be prepared by preparing a hybridoma through fusion of the antibody producing cells with myeloma cells of an animal and culturing the hybridoma, or administering the hybridoma to the animal to induce ascites tumor in the animal, and then isolating and purifying it from the culture medium or ascitic fluid. Also, a polyclonal antibody to the protein of the present invention can be prepared by isolating the immune serum of the immune animal. 
     Using the antibody of the present invention, the IgA nephropathy-related protein can be detected or determined immunologically. 
     Examples of the immunological detection method include ELISA method using a microtiter plate, fluorescent antibody technique, western blot technique, immunohistochemical staining and the like. 
     Examples of the immunological determination method include sandwich ELISA method in which, among antibodies which react with the protein of the present invention in solution, two monoclonal antibodies having different epitopes are used and radioimmunoassay method in which the protein of the present invention labeled with radioactive isotope, such as  125 I or the like, and an antibody which recognizes the protein of the present invention are used. 
     Using the antibody of the present invention, the presence or absence of IgA nephropathy in a person to be inspected can be diagnosed by immunologically detecting or determining an IgA nephropathy-related protein in leukocytes collected from a healthy person and the person to be inspected, comparing its amounts in the healthy person and person to be inspected and then examining the quantitative fluctuation. As a specific sample to be tested, leukocytes separated from peripheral blood samples of a healthy person and a person to be inspected can be used. Additionally, when the IgA nephropathy-related protein to be detected is a protein secreted from leukocytes, the presence or absence of IgA nephropathy in a person to be inspected can be detected and diagnosed by immunologically detecting or determining the protein in blood plasma samples collected from a healthy person and the person to be inspected, comparing its amounts in the healthy person and person to be inspected and then examining its quantitative fluctuation. 
     The antibody of the present invention can be applied to the treatment or prevention of IgA nephropathy. 
     When the DNA, protein, oligonucleotide and antibody is used for the diagnosis, treatment or prevention of IgA nephropathy, a diagnostically or pharmacologically acceptable carrier may be added. 
     EXAMPLES 
     Examples of the present invention are given below by way of illustration and not by way of limitation. 
     Example 1 
     Preparation of Human Undifferentiated Myeloid Cell Line KG-1cDNA Library 
     (1) Preparation of Poly (A) RNA from KG-1 Cell 
     KG-1 (ATCC CCL246), human undifferentiated myeloid cell line, was cultivated in the Iskov modified Dulbecco&#39;s medium (manufactured by Gibco BRL) to collect cells as pellet. To 1 ml of the cell pellet, 2.8 ml of a cytolytic solution [140 mM NaCl, 1.5 mM MgCl 2 , 10 mM tris(hydroximethyl)aminomethane (Tris)-HCl (pH 8.5), 0.5% of Nonidet P-40] and 0.2 ml of 400 mM vanadyl sulfuric acid ribonucleotide complex were added. The cells were dissolved in the solution on the ice, superposed on 1 ml of a saccharose solution [140 mM NaCl, 1.5 mM MgCl 2 , 10 mM Tris-HCl (pH 8.5), 1% Nonidet P-40, 24% saccharose], and centrifuged at 2° C. for 30 minutes at 30,000 rpm. To 4 ml of the supernatant containing RNA, 4 ml of an enzymatic reaction buffer [200 mM Tris-HCl (pH 7.5), 25 mM disodium ethylenediaminotetraacetate (EDTA), 300 mM NaCl, 2% lauryl sodium sulfate (SDS)] and 200 μl of 10 mg/ml proteinase K were added and allowed to react for 5 minutes at room temperature. To this reaction mixture, phenol extraction and chloroform extraction were carried out, and ethanol precipitation were carried out to recover RNA. After RNA was sufficiently dissolved in 4.5 ml of a buffer solution [20 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 0.1% SDS], it was heated at 70° C. for 5 minutes and chilled quickly, and then 0.5 ml of 5 M LiCl was added. The solution was centrifuged at 2° C. for 10 minutes at 8,000 rpm, the supernatant was heated at 70° C. for 5 minutes and chilled quickly, and then applied to an oligo d(T) cellulose column [Pasteur pipet charged with oligo dT cellulose (manufactured by Sigma)] which had been equilibrated with an absorption buffer [20 mM Tris-HCl (pH 7.5), 500 mM LiCl, 0.1 mM EDTA, 0.1% SDS]. The flow-through from the column was heated at 70° C. for 5 minutes and chilled quickly, and then applied to the column for absorbing poly (A)  + RNA again. The column was washed once with 1 ml of the absorption buffer, 3 times with 1.5 ml of the absorption buffer, 5 times with 1.5 ml of a washing buffer [20 mM Tris-HCl (pH 7.5) and 100 mM LiCl, 0.1 mM EDTA], and eluted with 3 ml of an elution buffer [10 mM Tris-HCl (pH 7.5), 0.1 mM EDTA] for recovering poly (A) + RNA. From 6 ml (2 g) of the cell pellet, 8 mg of total RNA was obtained, and 200 μg of poly (A) + RNA was obtained therefrom. 
     (2) Production of cDNA Library 
     A 8 μl (9.1 μg) portion of poly d(T)-NotI primer represented by SEQ ID NO:3, 32 μl of 5 mM Tris-HCl (pH 8.3) and 120 μl of distilled water were added to the obtained poly (A) + RNA 40 μg (40 μl). DNA was synthesized by the method described in  Nucleic Acids Res.,  12, 4539 (1984). It was heated at 70° C. for 10 minutes and chilled quickly, and then 62 μl of 5× reverse transcriptase reaction buffer [250 mM Tris-HCl (pH 8.3), 375 mM KCl, 15 mM MgCl 2 ], 32 μl of 100 ml dithiothreitol (DTT) and 8 μl of 20 mM dNTP (dATP, dGTP, dTTP, dCTP) were added thereto. After heating at 37° C. for 2 minutes, 3,200 units (16 μl) of reverse transcriptase SUPERSCRIPT RNase H −  Reverse Transcriptase (manufactured by Gibco BRL) were added thereto, and the solution was allowed to react at 37° C. for 1 hour to synthesize cDNA. To the reaction mixture, 15 μl of 20 mM dNTP, 60 μl of 100 mM DTT, 16 μl of 15 mM β-nicotinamide dinucleotide and 929 μl of distilled water were added. Thereto further added was 32 units (16 μl) of  E. coli  ribonuclease H (manufactured by Takara Shuzo Co., Ltd.), 256 units (32 μl) of DNA polymerase I (manufactured by Takara Shuzo Co., Ltd.) and 480 units (8 μl) of  E. coli  DNA ligase (manufactured by Takara Shuzo Co., Ltd.), and allowed to react at 16° C. for 2 hours to prepare cDNA double-stranded. T4 DNA polymerase (48 units (24 μl)) was added and allowed to react at 16° C. for 5 minute to convert the termini blunt. EDTA (24 μl, 200 mM) was added thereto, and heated at 70° C. for 10 minutes to stop the reaction. The reaction mixture was subjected to phenol-chloroform extraction, and then ethanol precipitation was carried out. Additionally, ethanol precipitation was carried out two times to purify the cDNA. The double-stranded cDNA was dissolved in 40 μl of TE buffer [10 mM Tris-HCl (pH 8.0), 1 mM EDTA (pH 8.0)], and 60 μl of 10× restriction enzyme reaction buffer [500 mM Tris-HCl (pH 7.5), 100 mM MgCl 2 , 10 mM DTT, 1M NaCl, 0.1% Triton X-100, 0.1% bovine serum albumin (BSA)] and 460 μl of distilled water were added thereto, and 320 units (40 μl) of restrictive enzyme NotI (manufactured by Takara Shuzo Co., Ltd.) were added thereto for digestion reaction at 37° C. for 2 hours. Moreover, 192 units (24 μl) of NotI were added thereto, and the reaction was carried out at 37° C. for 1 hour, and the reaction mixture was heated at 70° C. for 3 minutes to stop the reaction. After phenol/chloroform extraction, ethanol precipitation was carried out, and additionally, ehtanol precipitation was carried out two time to purify cDNA. The cDNA was centrifuged at 24,000 rpm for 24 hours with sucrose density gradient of 5 to 30% to separate into the fraction of larger than 6 kb, the fraction of 2 to 6 kb and the fraction of less than 2 kb. The fraction of 2 to 6 kb (0.5 μg) and 0.5 μg of plasmid vector pBluescript SK+ digested by NotI-EcoRV are ligated and introduced into  E. coli  DH10B (manufactured by Gibco) to prepare the cDNA library. 
     (3) Determination of Nucleotide Sequence of cDNA of Each Clone in KG-1cDNA Library 
     The nucleotide sequence of cDNA of each clone in the library was determined using 373 DNA sequencer of Parkin Elmer. As specific reagents and method for the nucleotide sequence determination, Dye Primer Cycle Sequencing FS Ready Reaction Kit of Parkin Elmer were used in accordance with the manual attached to the kit. Thus obtained cDNA nucleotide sequences were collected to create a database. 
     Example 2 
     Cloning of cDNA Fragment of the Novel Gene KIAA0235 
     (1) Preparation of Total RNA from Leukocytes of IgA Nephropathy Patients and Healthy Persons 
     A 20 ml portion of blood was collected from each of five IgA nephropathy patients and five healthy persons. Each blood sample was mixed with 500 μl of 1,000 units/ml heparin solution to inhibit coagulation, transferred into a centrifugation tube and then centrifuged at 3,300 rpm for 15 minutes at room temperature, and the resulting intermediate layer buffy coat containing leukocytes was transferred into another centrifugation tube. Thereafter, total RNAs were obtained in accordance with the AGPC method [Experimental Medicine, 9 , 1937 (1991)]. 
     (2) Fluorescence Differential Display Using Leukocyte Total RNAs of IgA Nephropathy Patients and Healthy Persons 
     Distilled water was added to 2.5 μg of each of the total RNAs to a total volume of 9 μl, and the solution was mixed with 1 μl of an anchor primer (nucleotide sequence represented by SEQ ID NO:4, custom-synthesized by Sawady, 50 μM) whose 5′-end had been fluorescence-labeled with fluorescein isothiocyanate (referred to as “FITC” hereinafter), heated at 70° C. for 5 minutes and then immediately cooled on an ice bath. Thereto added were 2 μl of distilled water, 4 μl of 5× reverse transcriptase reaction buffer, 2 μl of 100 mM DTT, 1 μl of 10 mM dNTP and 1 μl (200 units) of reverse transcriptase SUPERSCRIPT RNase H −  Reverse Transcriptase (manufactured by Gibco BRL), the solution was mixed, allowed to react at 25° C. for 10 minutes and at 42° C. for 50 minutes to synthesize cDNA, and heated at 90° C. for 5 minutes to stop the reaction. To the reaction mixture, 80 μl of TE buffer was added. 
     To 1 μl of each synthesized cDNA, 14.7 μl of distilled water, 2 μl of 10×PCR buffer [100 mM Tris-HCl (pH 8.8), 500 mM KCl, 15 mM MgCl 2 , 1% Triton X-100], 0.8 μl of 2.5 mM dNTP, 0.3 μl of 50 μM fluorescent labeling anchor primer FCH, 1 μl of arbitrary primer OPD-16 (nucleotide sequence represented by SEQ ID NO:5, manufactured by Operon, 10 μM) and 0.2 μl of DNA polymerase Gene Taq (manufactured by Nippon Gene Co., Ltd. 5 units/μl) were added and set to a thermal cycler. The PCR was effected by carrying out the reaction at 94° C. for 3 minutes, 40° C. for 5 minutes and 72° C. for 5 minutes, subsequently carrying out a total of 27 cycles of the reaction in which one cycle was composed of the steps of 95° C. for 15 seconds, 40° C. for 2 minutes and 72° C. for 1 minute, and finally carrying out 5 minutes of the reaction at 72° C. 
     A 3 μl portion of an electrophoresis sample solution (95% formaldehyde, 0.1% xylenecyanol, 0.1% bromphenol blue) to 4 μl of each PCR mixture, heated at 95° C. for 2 minutes and chilled immediately, and 6% acrylamide gel electrophoresis was conducted at and 1500 V for 2.5 hours. The electrophoresis buffer containing 89 mM Tris, 89 mM boric acid, 2 mM EDTA were used. Gel fluorescence after the electrophoresis was measured by a fluorimager (manufactured by Molecular Dynamics Inc.) for detecting and comparing the PCR amplified fragments. In comparison with 5 cases of the healthy persons, a band which significantly increased in leukocytes of 5 cases of the IgA nephropathy patients was recorded. Furthermore, total RNAs were prepared from other 3 cases of IgA nephropathy patients and 3 cases of healthy persons in the same manner to carry out the differential display in the same manner. 
     A band of about 200 bands which showed increased fluorescence in both of the above two trials of the differential display was cut off from the gel. 
     A 38 μl portion of distilled water, 5 μl of 10×PCR buffer, 4 μl of 2.5 mM dNTP, non-fluorescence labeling anchor primer NC (sequence listing represented by SEQ ID NO:4, produced by Sawady, 34 μm), 2 μl of 10 μM arbitrary primer OPD-16 (sequence listing represented by SEQ ID NO:5) and 0.5 μl of DNA polymerase Gene Taq were added to about ¼ portion of the gel thus cut off, the resulting mixture was heated at 94° C. for 3 minutes and then a total of 30 cycles of the reaction was carried out in which one cycle was comprised of the steps of 95° C. for 15 seconds, 40° C. for 2 minutes and 72° C. for 1 minute, subsequently carrying out 5 minutes of the reaction at 72° C. to complete PCR. 
     The resulting reaction solution was extracted with phenol-chloroform (1:1) and then with chloroformisoamyl alcohol (24:1), subsequently carrying out ethanol precipitation. The resulting precipitation was dissolved in 10 μl of TE buffer. 
     A 1 μl portion of amplified fragment and 1 μl of PCR fragment cloning vector pT7BlueT-Vector (manufactured by Novagen Inc.) were mixed and the amplified fragment was inserted into the plasmid using DNA ligation kit ver. 1 (manufactured by Takara Shuzo Co., Ltd.) according to the instructions of the kit.  E. coli  DH5α (manufactured by Gibco BRL) was transformed with the ligated mixture to obtain an ampicillin resistant transformant. Plasmid DNA was isolated for the transformant strain according to a publicly known method. A 0.3 ng portion of the plasmid DNA was dissolved into 19 μl of distilled water, and 2.5 μl of 10×PCR buffer, 2 μl of 2.5 mM dNTP, 0.3 μl of 34 μM anchor primer NC, 1 μl of 10 μM optional primer OPD-16 and 0.5 μl of DNA polymerase Gene Taq were added for performing PCR under the same conditions reamplification of amplified fragment. Since the fragment of about 200 bp was amplified, it was confirmed that the amplified fragment had been inserted into the plasmid. 
     (3) Determination of Amplified Fragment Nucleotide Sequence 
     The nucleotide sequence of the amplified fragment was determined by means of DNA sequencer (manufactured by Parkin Elmer). For the nucleotide sequence determination, reagents and method supplied by Dye Primer Cycle Sequencing FS Ready Reaction Kit of Parkin Elmer were used according to the instructions of the kit. The thus obtained nucleotide sequence corresponded to the DNA sequence 1076 to 1261 in SEQ ID NO:1. Thus obtained nucleotide sequence was compared with the nucleotide sequence database GenBank. The nucleotide sequence corresponding to the concerned nucleotide sequence was not identified among nucleotide sequences in the database, so that it was considered as cDNA fragment of a novel gene. Then the nucleotide sequence was compared with cDNA nucleotide sequence data base in the KG-1cDNA library, it was found to completely agree with a portion of cDNA nucleotide sequence of the gene KIAA0235, determining that KIAA0235 was a novel gene whose expression increases in the leukocyte of IgA nephropathy patients. As shown in SEQ ID NO:1, KIAA0235 was 5399 bp long and an open reading frame (ORF) composed of 850 amino acids exist therein. The amino acid sequence thereof is shown in SEQ ID NO:2. In the comparison of KIAA0235 nucleotide sequence with the database Genbank/EMBL, there are no identical nucleotide sequence, confirming that it concerns a novel gene cDNA. Moreover, when the amino acid sequence encoded by the cDNA ORF was compared with the database, it has shown a homology, at C terminal side, with the amino acid sequence of the pumilio gene [ Development,  114, 221 (1992);  Cell,  80, 747 (1995)] that is bound with mRNA of the hunchback gene derived from the mother, during the embryo development of Drosophila, and playing an important role in the formation of abdominal arthromere. It has also shown a homology of 82% with the amino acid sequence of the ORF of KIAA0099 [ DNA Research,  2, 37 (1995)] already reported in the cDNA clone derived from this KG-1. 
     Example 3 
     Northern Blot of KIAA0235 in Human Cell and Various Tissues 
     Next, Northern blot was carried out to determine the expression of mRNA of the novel gene KIAA0235 in human cell and tissues. 
     Using filters manufactured by Clonetech blotting mRNA of human various tissues, the expression of mRNA of KIAA0235 in human various tissues was detected by means of Northern blot. Heart, brain, placenta, lung, lever, skeletal muscle, kidney and pancreas mRNAs were blotted on the filter, Human MTN blot. Spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral leukocyte mRNAs were blotted in the Human MTN Blot II. Concerning the cell, 2 μg of poly (A)+RNA obtained from human cervical carcinoma cell line, HeLa cell in the same manner as in Example 1 and poly (A)  + RNA of KG-1 were applied to agarose gel electrophoresis, transferred to filter Biodyne A (manufactured by Paul Inc.), and KIAA0235 mRNA expression was detected by means of Northern blotting in the same manner. To use as probe, plasmid clone HA4677 containing KIAA0235 cDNA in the cDNA library prepared in Example 1 was cleaved by NotI (manufactured by Takara Shuzo Co., Ltd.) and HindIII (manufactured by Takara Shuzo Co., Ltd.) to purify KIAA0235 cDNA fragment. A 50 ng portion of this fragment were  32 P labeled using [α- 32 P]dCTP and BcaBEST labeling kit (manufactured by Takara Shuzo Co., Ltd.). Specific reagents and method of the labeling were complied with the instructions of the kit. 
     The filter was soaked in a hybridization solution (50% formaldehyde, 5×SSC, 5× Denhardt&#39;s solution (0.1% BSA, 0.1% ficoal, 0.1% polyvinyl pyrolidone, 0.25% SDS, 100 μg/ml of herring spermatozoa DNA which had been subjected to ultrasonic treatment and then denaturation (the process of seething in a microwave oven and chilled quickly in the ice was repeated 3 times), sealed and prehybridized overnight at 37° C. Next, the probe was denatured by seething in a microwave oven and chilling in the ice 3 times, added into the hybridization solution. The filter was soaked therein, sealed and hybridized overnight at 37° C. 
     The filter was taken out, washed by shaking in 1×SSC containing 0.1% SDS at room temperature. Changing the solution, it was washed furthermore several times, then rinsed with 0.1×SSC containing 0.5 SDS, and washed by shaking at 50° C. for 1 hour in 0.1×SSC containing 0.5% SDS. Finally, it was washed with 1×SSC containing 0.1% SDS and dried in the air. The filter was overlaid on the imaging plate (manufactured by Fuji Photo Film Co., Ltd.) for conducting the autoradiography for 4 hours, and analyzed with Bioimaging Analyzer BAS 2000 (manufactured by Fuji Photo Film Co. Ltd.). The results are shown in FIG.  1 . Bands corresponding to mRNA of 2 different lengths, 5.5 kb and 6.8 kb, were detected in all tissues of heart, brain, placenta, lung, lever, skeletal muscle, kidney, pancreas, spleen, thymus, prostate, testis, ovary, small intestine, colon and peripheral leukocyte. Therefore, it was confirmed that KIAA0235 was a gene expressing ubiquitously in respective tissue. Moreover, bands corresponding to mRNA of 2 different lengths, 5.5 kb and 6.8 kb, were similarly detected in KG-1 and HeLa cells, and the expression of KIAA0235 in both cells was confirmed. 
     Example 4 
     Detection of Expression Specificity of KIAA0235 by RT-PCR 
     Using single-stranded cDNA synthesis kit, Superscript Preamplification System (manufactured by BRLs), to 2 μg of total RNA from leukocyte of 5 IgA nephropathy patients and 5 healthy persons obtained in Example 2, single-stranded cDNA was synthesized by oligo dT primer supplied by the kit. Specific reagents and method were complied with the protocol attached to the kit. After the reaction, 399 μl of distilled water was added to 21 μl of reaction mixture to be 420 μl as a whole, mRNA expression amount corresponding to each amplified fragment was measured by using 10 μl thereof according to RT-PCR. Namely, 15.8 μl of distilled water, 4 μl of 10×PCR buffer, 3.2 μl of 2.5 mM dNTP, 2 μl of DMSO, 2 μl of 10 μM KIAA0235 specific 5′-side sense primer (SEQ ID NO:6), 2 μl of 10 μM KIAA0235 specific 3′-side antisense primer (SEQ ID NO:7) and 2 μl of DNA polymerase GeneTag diluted to 1 unit/μl were added to 10 μl of leukocyte single-stranded cDNA, heated at 97° C. for 5 minutes, chilled quickly in the ice for 5 minutes. Subsequently, the PCR (a total of 30 cycles of the reaction in which one cycle was composed of the steps of 95° C. for 30 seconds, 65° C. for 1 minute and 72° C. for 2 minutes) was carried out. 
     In order to make a correction of the mRNA amount, with regard to glyceraldehyde-3-phosphate dehydrogenase (G3PDH) gene, a housekeeping gene, the similar reaction was carried out using a specific primer represented by SEQ ID NO:8 and NO:9, the mRNA expression amount of each gene was calibrated by the ratio of the expression amount of G3PDH mRNA, and then the average values of 5 IgA nephropathy patients and 5 healthy persons were compared. The results are shown in FIG.  2 . It was confirmed that in the leukocyte of IgA nephropathy patients, KIAA0235 has expressed 6.6 times more than in healthy persons. 
     While the invention has been described in detail and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that various changes and modifications can be made therein without departing from the spirit and scope thereof. 
     
       
         
           
             9 
           
           
             1 
             5399 
             DNA 
             Homo sapiens 
             
               CDS 
               (2)..(2551) 
               Cell Line KG-1 
             
           
            1
a gaa ttt tca aat cct gaa act cag aat ctg gat gcc atg gaa caa gtt     49
  Glu Phe Ser Asn Pro Glu Thr Gln Asn Leu Asp Ala Met Glu Gln Val
    1               5                  10                  15
ggt ctg gaa tcc tta cag ttt gac tat cct ggt aat cag gta cca atg       97
Gly Leu Glu Ser Leu Gln Phe Asp Tyr Pro Gly Asn Gln Val Pro Met
             20                  25                  30
gac tct tca gga gct act gta ggc ctt ttt gac tac aat tcc cag cag      145
Asp Ser Ser Gly Ala Thr Val Gly Leu Phe Asp Tyr Asn Ser Gln Gln
         35                  40                  45
cag ctc ttt cag agg act aat gca cta aca gtt caa cag tta act gca      193
Gln Leu Phe Gln Arg Thr Asn Ala Leu Thr Val Gln Gln Leu Thr Ala
     50                  55                  60
gct caa cag cag caa tat gca tta gca gca gct cag cag cca cat ata      241
Ala Gln Gln Gln Gln Tyr Ala Leu Ala Ala Ala Gln Gln Pro His Ile
 65                  70                  75                  80
gct ggt gta ttc tca gca ggc ctt gct cca gct gca ttt gtg cca aat      289
Ala Gly Val Phe Ser Ala Gly Leu Ala Pro Ala Ala Phe Val Pro Asn
                 85                  90                  95
cca tac att att agt gct gct cct cca ggg acc gat ccg tat act gca      337
Pro Tyr Ile Ile Ser Ala Ala Pro Pro Gly Thr Asp Pro Tyr Thr Ala
            100                 105                 110
gca gga ttg gct gca gca gct aca tta gca ggt cca gca gtg gtt cca      385
Ala Gly Leu Ala Ala Ala Ala Thr Leu Ala Gly Pro Ala Val Val Pro
        115                 120                 125
cct cag tat tac ggc gtt cca tgg ggg gtg tat cca gcc aac tta ttt      433
Pro Gln Tyr Tyr Gly Val Pro Trp Gly Val Tyr Pro Ala Asn Leu Phe
    130                 135                 140
cag cag caa gct gca gct gcg gca aat aac aca gcc agt cag caa gca      481
Gln Gln Gln Ala Ala Ala Ala Ala Asn Asn Thr Ala Ser Gln Gln Ala
145                 150                 155                 160
gca tca caa gct cag cct gga cag caa cag gtt ctc cgt gct gga gca      529
Ala Ser Gln Ala Gln Pro Gly Gln Gln Gln Val Leu Arg Ala Gly Ala
                165                 170                 175
ggt cag cgt cct ctt act ccc aat cag ggt cag caa ggg cag caa gca      577
Gly Gln Arg Pro Leu Thr Pro Asn Gln Gly Gln Gln Gly Gln Gln Ala
            180                 185                 190
gaa tca ctt gcg gca gct gca gca gca aat cca aca ttg gct ttt ggt      625
Glu Ser Leu Ala Ala Ala Ala Ala Ala Asn Pro Thr Leu Ala Phe Gly
        195                 200                 205
cag ggt ctt gct act ggc atg cca ggc tat caa gta cta gct cca act      673
Gln Gly Leu Ala Thr Gly Met Pro Gly Tyr Gln Val Leu Ala Pro Thr
    210                 215                 220
gcc tat tat gat cag act ggt gcc tta gtg gtt ggc cct gga gca agg      721
Ala Tyr Tyr Asp Gln Thr Gly Ala Leu Val Val Gly Pro Gly Ala Arg
225                 230                 235                 240
act ggc ctt gga gct cca gtt cgg tta atg gct cca aca cct gtt tta      769
Thr Gly Leu Gly Ala Pro Val Arg Leu Met Ala Pro Thr Pro Val Leu
                245                 250                 255
att agt tca gca gca gca caa gct gca gca gca gca gca gct gga gga      817
Ile Ser Ser Ala Ala Ala Gln Ala Ala Ala Ala Ala Ala Ala Gly Gly
            260                 265                 270
act gca agt agc ctt aca ggc agc aca aat ggt ctg ttt cgg cca att      865
Thr Ala Ser Ser Leu Thr Gly Ser Thr Asn Gly Leu Phe Arg Pro Ile
        275                 280                 285
ggc act cag cca cca cag cag cag caa cag cag cca agc act aat ctg      913
Gly Thr Gln Pro Pro Gln Gln Gln Gln Gln Gln Pro Ser Thr Asn Leu
    290                 295                 300
caa tct aat tca ttt tat gga agc agt tct ttg act aat agc tcc cag      961
Gln Ser Asn Ser Phe Tyr Gly Ser Ser Ser Leu Thr Asn Ser Ser Gln
305                 310                 315                 320
agt agt tct tta ttt tct cat gga cct ggt caa cct gga agt aca tct     1009
Ser Ser Ser Leu Phe Ser His Gly Pro Gly Gln Pro Gly Ser Thr Ser
                325                 330                 335
ctt ggc ttt gga agt ggt aac tct ttg ggt gct gct ata ggc tca gcc     1057
Leu Gly Phe Gly Ser Gly Asn Ser Leu Gly Ala Ala Ile Gly Ser Ala
            340                 345                 350
ctc agt gga ttt ggt tca tca gtt ggc agt tct gca agt agt agt gcc     1105
Leu Ser Gly Phe Gly Ser Ser Val Gly Ser Ser Ala Ser Ser Ser Ala
        355                 360                 365
aca agg aga gag tct cta tct act agc tct gac ttg tac aaa aga tct     1153
Thr Arg Arg Glu Ser Leu Ser Thr Ser Ser Asp Leu Tyr Lys Arg Ser
    370                 375                 380
agt agc agc cta gca ccc ata ggg caa cca ttt tac aat agt ctg gga     1201
Ser Ser Ser Leu Ala Pro Ile Gly Gln Pro Phe Tyr Asn Ser Leu Gly
385                 390                 395                 400
ttt tcc tcc tct cca agt cca ata ggc atg cct ctg cca agc caa act     1249
Phe Ser Ser Ser Pro Ser Pro Ile Gly Met Pro Leu Pro Ser Gln Thr
                405                 410                 415
cca gga cat tca ctt acg cca ccg cca tca ctt tca tca cat gga tcc     1297
Pro Gly His Ser Leu Thr Pro Pro Pro Ser Leu Ser Ser His Gly Ser
            420                 425                 430
tca tcc agt ttg cat tta gga gga ctg aca aat ggt agt ggt cga tat     1345
Ser Ser Ser Leu His Leu Gly Gly Leu Thr Asn Gly Ser Gly Arg Tyr
        435                 440                 445
atc tct gca gca cct gga gca gaa gca aaa tat cga agt gct tca agc     1393
Ile Ser Ala Ala Pro Gly Ala Glu Ala Lys Tyr Arg Ser Ala Ser Ser
    450                 455                 460
act tcc agt cta ttt agc tcc agc agc cag ctc ttt cct cct tcc cgg     1441
Thr Ser Ser Leu Phe Ser Ser Ser Ser Gln Leu Phe Pro Pro Ser Arg
465                 470                 475                 480
ctt cgg tat aat agg tct gat att atg cct tct ggc cgc agt aga tta     1489
Leu Arg Tyr Asn Arg Ser Asp Ile Met Pro Ser Gly Arg Ser Arg Leu
                485                 490                 495
ttg gaa gat ttc aga aac aac cgc ttc cca aac ctt cag ctt aga gac     1537
Leu Glu Asp Phe Arg Asn Asn Arg Phe Pro Asn Leu Gln Leu Arg Asp
            500                 505                 510
ttg att gga cat ata gtt gag ttt tct caa gac cag cat ggt tct aga     1585
Leu Ile Gly His Ile Val Glu Phe Ser Gln Asp Gln His Gly Ser Arg
        515                 520                 525
ttc ata cag caa aaa cta gag aga gct act cca gct gag cga cag atg     1633
Phe Ile Gln Gln Lys Leu Glu Arg Ala Thr Pro Ala Glu Arg Gln Met
    530                 535                 540
gta ttt aat gaa att ctg caa gca gcc tat caa tta atg act gat gtt     1681
Val Phe Asn Glu Ile Leu Gln Ala Ala Tyr Gln Leu Met Thr Asp Val
545                 550                 555                 560
ttt ggc aac tat gtt ata cag aag ttt ttt gag ttt ggg agt ctg gat     1729
Phe Gly Asn Tyr Val Ile Gln Lys Phe Phe Glu Phe Gly Ser Leu Asp
                565                 570                 575
caa aaa tta gcc ctg gct act cgt att cgt ggt cat gtt cta ccc tta     1777
Gln Lys Leu Ala Leu Ala Thr Arg Ile Arg Gly His Val Leu Pro Leu
            580                 585                 590
gcc ttg cag atg tat ggc tgc cgc gtt att cag aaa gca tta gaa tct     1825
Ala Leu Gln Met Tyr Gly Cys Arg Val Ile Gln Lys Ala Leu Glu Ser
        595                 600                 605
att tct tct gac cag cag agt gaa atg gta aag gag ctg gat ggt cat     1873
Ile Ser Ser Asp Gln Gln Ser Glu Met Val Lys Glu Leu Asp Gly His
    610                 615                 620
gtg ctc aaa tgt gtg aaa gat cag aat gga aac cat gtt gta caa aaa     1921
Val Leu Lys Cys Val Lys Asp Gln Asn Gly Asn His Val Val Gln Lys
625                 630                 635                 640
tgt atc gaa tgt gtt cag cca cag tca cta cag ttc atc att gat gct     1969
Cys Ile Glu Cys Val Gln Pro Gln Ser Leu Gln Phe Ile Ile Asp Ala
                645                 650                 655
ttc aag gga caa gta ttt gtg ctt tca act cat cct tat ggc tgc aga     2017
Phe Lys Gly Gln Val Phe Val Leu Ser Thr His Pro Tyr Gly Cys Arg
            660                 665                 670
gta att cag cgc atc cta gag cat tgc act gca gaa cag acc tta cct     2065
Val Ile Gln Arg Ile Leu Glu His Cys Thr Ala Glu Gln Thr Leu Pro
        675                 680                 685
atc tta gaa gaa ctc cac caa cat aca gag cag ttg gta cag gat cag     2113
Ile Leu Glu Glu Leu His Gln His Thr Glu Gln Leu Val Gln Asp Gln
    690                 695                 700
tat ggc aat tat gtt att cag cat gta ctg gaa cac ggt cga cct gaa     2161
Tyr Gly Asn Tyr Val Ile Gln His Val Leu Glu His Gly Arg Pro Glu
705                 710                 715                 720
gac aag agc aaa att gtt tcc gaa atc agg gga aag gtt tta gcc ctg     2209
Asp Lys Ser Lys Ile Val Ser Glu Ile Arg Gly Lys Val Leu Ala Leu
                725                 730                 735
agt caa cac aaa ttt gcc agc aat gta gta gaa aag tgt gtt act cat     2257
Ser Gln His Lys Phe Ala Ser Asn Val Val Glu Lys Cys Val Thr His
            740                 745                 750
gcc tcc cgt gct gag aga gct tta ctg att gac gag gtt tgc tgc cag     2305
Ala Ser Arg Ala Glu Arg Ala Leu Leu Ile Asp Glu Val Cys Cys Gln
        755                 760                 765
aat gat ggt cct cac agt gcc tta tac acc atg atg aag gac cag tat     2353
Asn Asp Gly Pro His Ser Ala Leu Tyr Thr Met Met Lys Asp Gln Tyr
    770                 775                 780
gcc aat tac gtg gtt caa aag atg att gat atg gct gaa cct gct cag     2401
Ala Asn Tyr Val Val Gln Lys Met Ile Asp Met Ala Glu Pro Ala Gln
785                 790                 795                 800
aga aag ata atc atg cac aag att cga cct cac att act act ttg cgc     2449
Arg Lys Ile Ile Met His Lys Ile Arg Pro His Ile Thr Thr Leu Arg
                805                 810                 815
aaa tac aca tac ggg aag cat ata ctg gcc aag ttg gaa aag tat tat     2497
Lys Tyr Thr Tyr Gly Lys His Ile Leu Ala Lys Leu Glu Lys Tyr Tyr
            820                 825                 830
ttg aag aat agc ccg gac cta gga cct att gga gga cca cca aat gga     2545
Leu Lys Asn Ser Pro Asp Leu Gly Pro Ile Gly Gly Pro Pro Asn Gly
        835                 840                 845
atg ctg taaattacag gagcaagaga aagaagataa tttaaccatg tgaaaagaat      2601
Met Leu
    850
ttttttgtgc gtgaattatc aaaacacaac tcaactatga atcttcaatt tttttttaaa   2661
gcaaaactat ttattgactt tattcatcca tttgtaaatt ttttaaggtt cttgtgtata   2721
tttggggggt gggggatgaa ttataaatta tattcagccc tgagtggaga cctatcagat   2781
tggattgctg gcaaagcaca gaatgcctgt atatgatgta actgtatcaa aaataaaaag   2841
ctgtcacata ttttgtaaat ttttaccttg taaagtcaca aaaatagttt ttaaaggaaa   2901
aagtacagta ttcttttaat aaactggctc acagtctggt aggtctacaa ccccatagca   2961
caacaggttt atagagatgt atatagaatt atagtcctta tttttttcct ttgcgtgaaa   3021
ccttttataa cagattaaca atcaactgca taaatattat taatatttta aaaagagtta   3081
agttgtattt tgataattca caaactatca tgcaaataac gagtaagtag acaagaataa   3141
agtggtttga gatgaaaaga acctaacatt atttacagta gatgtggttt taatacaatt   3201
actgccctaa aatgtctctg gcaatgtaca gaaatattgt atatacttac atatgtaatt   3261
gttgtaagag ttaaatacaa aatcatggtg acacttccaa ttaagtgcac taaatgaaaa   3321
gttaagtcac ttattaactt ttcagtttgg tttgcaatga gaaagagtgg aaatttgtat   3381
tttgttttgc ttatagaatt acagacatgt tgaggaagtg ttgagcttta ttttgctttt   3441
tcatagaggc agaaagtagg aaccagatag agatgaaaag gggccactga aaagtgaatt   3501
tgatagctca gcatttaagc atgattacat attcagatag ctctttttgc tttctataaa   3561
tatatgcatt gtgtgtgtag taatagatgt aagtttacac tttgaaagga aatcttgttt   3621
caatgtttat tataaaagcc ttgctaattt agtagtgatg ctttccttgg ttgtacaggt   3681
gtacatttgt aaaccttcat gctgtaaatg gaatttgttt tatctctttg ggatacattt   3741
gcattttagt gtacatttac gtccctgccc tctttgacct ggcaatatag tgttgtataa   3801
tgtaaattta tttctccaaa tcgagagtga ttttttaaaa attttttatc tttatatggt   3861
ttcagaagta tgaaccagct ttctttttat tattgtgaga tcattttgtt ttataacata   3921
gttgttgact gttaatatgg acctgctaga atttggatca ctttcaattg aagtcagggt   3981
attgtgcata atagaaagta ttggactgag atatttggtt accatggagg ccaatgcttt   4041
tttcatctta ttaaatgtga tgtgactttt ttctttgtac agaagagtac tgtatttttg   4101
aatagcctac tcccaagtaa gagcaaatct gtatgataac attttttcct ctggacataa   4161
gacataacag taacacgatg tacatttaca agcggcctta tgtacatttc ccaacaatct   4221
ttttaaggca aaattgtgac catatgtgta taattaaaat cgtttttaat cctttgccta   4281
tgaaaatatt ttggaaaaaa acttgctgtg tatattcagt ttctgaaaga taaagaaagt   4341
gctttgtatt ttgttgaagt cagtattttg tataaacatt tatgttgacc cacttatgtt   4401
cagtgctgaa aactaaaatg aacatgctat tctgtcagct gaatatggaa gagatctttt   4461
tttactagag atctgcagaa gaaacgcaat cttctgagca caatatggaa tctaaaggtt   4521
ttatcactta gttgttcata ttatgaacct aaaaataatg gcataaagtt tggggatgcc   4581
aggcatactt tttcatgttt ggtgttgagt tattttactt ttctaaccca acattccttg   4641
gtgagaccat taaatccaaa cacttgtcac cgttccttct catagtcact ctgggtcatc   4701
agcatgtccc agtcactgca gcaacgcctt gtgtttgttt cattttttta aaacccacac   4761
aaagccgctg tctcactttt tcctacttta ccaacctcag agtatttcgg cccgtatcga   4821
acttttgttc tcagtatcag cccatggttt caggatcaaa gctgtcatgt tggagattgg   4881
taatggcttt cctgtctttg tacagttgaa ttcctagtct tccttcatcc ttgccctctg   4941
ttggcacagg cattatctct gcaattttag aaaatgacaa gtagagaata ctacattgag   5001
aaactaaacc ctcttcttgg ggtcctgata ctcattccca tttgtcccag tgctgacaac   5061
ccaatcttcc caatactttc aggcctgctc tacaaaagta cctgttcttg tagaaatttt   5121
acagtctgcc attttgggtg cccaccccaa tttttacctt ttagtaagtt ggcatgaaat   5181
tttggtaaaa tctgaaaatc acatttcaga ataaaacaat tgggcaaaac tacctaggct   5241
ttactcttga gtgtctcctt ttgataggga ttgtttctgg accagtttgt ctaagtcctg   5301
gctcttattg gttcatatga aataatgtta acttcacttc tttgtatatt atgtataaat   5361
tagaaaatga aaaatgtgtg aataacattg tatgaaat                           5399
 
           
             2 
             850 
             PRT 
             Homo sapiens 
           
            2
Glu Phe Ser Asn Pro Glu Thr Gln Asn Leu Asp Ala Met Glu Gln Val
  1               5                  10                  15
Gly Leu Glu Ser Leu Gln Phe Asp Tyr Pro Gly Asn Gln Val Pro Met
             20                  25                  30
Asp Ser Ser Gly Ala Thr Val Gly Leu Phe Asp Tyr Asn Ser Gln Gln
         35                  40                  45
Gln Leu Phe Gln Arg Thr Asn Ala Leu Thr Val Gln Gln Leu Thr Ala
     50                  55                  60
Ala Gln Gln Gln Gln Tyr Ala Leu Ala Ala Ala Gln Gln Pro His Ile
 65                  70                  75                  80
Ala Gly Val Phe Ser Ala Gly Leu Ala Pro Ala Ala Phe Val Pro Asn
                 85                  90                  95
Pro Tyr Ile Ile Ser Ala Ala Pro Pro Gly Thr Asp Pro Tyr Thr Ala
            100                 105                 110
Ala Gly Leu Ala Ala Ala Ala Thr Leu Ala Gly Pro Ala Val Val Pro
        115                 120                 125
Pro Gln Tyr Tyr Gly Val Pro Trp Gly Val Tyr Pro Ala Asn Leu Phe
    130                 135                 140
Gln Gln Gln Ala Ala Ala Ala Ala Asn Asn Thr Ala Ser Gln Gln Ala
145                 150                 155                 160
Ala Ser Gln Ala Gln Pro Gly Gln Gln Gln Val Leu Arg Ala Gly Ala
                165                 170                 175
Gly Gln Arg Pro Leu Thr Pro Asn Gln Gly Gln Gln Gly Gln Gln Ala
            180                 185                 190
Glu Ser Leu Ala Ala Ala Ala Ala Ala Asn Pro Thr Leu Ala Phe Gly
        195                 200                 205
Gln Gly Leu Ala Thr Gly Met Pro Gly Tyr Gln Val Leu Ala Pro Thr
    210                 215                 220
Ala Tyr Tyr Asp Gln Thr Gly Ala Leu Val Val Gly Pro Gly Ala Arg
225                 230                 235                 240
Thr Gly Leu Gly Ala Pro Val Arg Leu Met Ala Pro Thr Pro Val Leu
                245                 250                 255
Ile Ser Ser Ala Ala Ala Gln Ala Ala Ala Ala Ala Ala Ala Gly Gly
            260                 265                 270
Thr Ala Ser Ser Leu Thr Gly Ser Thr Asn Gly Leu Phe Arg Pro Ile
        275                 280                 285
Gly Thr Gln Pro Pro Gln Gln Gln Gln Gln Gln Pro Ser Thr Asn Leu
    290                 295                 300
Gln Ser Asn Ser Phe Tyr Gly Ser Ser Ser Leu Thr Asn Ser Ser Gln
305                 310                 315                 320
Ser Ser Ser Leu Phe Ser His Gly Pro Gly Gln Pro Gly Ser Thr Ser
                325                 330                 335
Leu Gly Phe Gly Ser Gly Asn Ser Leu Gly Ala Ala Ile Gly Ser Ala
            340                 345                 350
Leu Ser Gly Phe Gly Ser Ser Val Gly Ser Ser Ala Ser Ser Ser Ala
        355                 360                 365
Thr Arg Arg Glu Ser Leu Ser Thr Ser Ser Asp Leu Tyr Lys Arg Ser
    370                 375                 380
Ser Ser Ser Leu Ala Pro Ile Gly Gln Pro Phe Tyr Asn Ser Leu Gly
385                 390                 395                 400
Phe Ser Ser Ser Pro Ser Pro Ile Gly Met Pro Leu Pro Ser Gln Thr
                405                 410                 415
Pro Gly His Ser Leu Thr Pro Pro Pro Ser Leu Ser Ser His Gly Ser
            420                 425                 430
Ser Ser Ser Leu His Leu Gly Gly Leu Thr Asn Gly Ser Gly Arg Tyr
        435                 440                 445
Ile Ser Ala Ala Pro Gly Ala Glu Ala Lys Tyr Arg Ser Ala Ser Ser
    450                 455                 460
Thr Ser Ser Leu Phe Ser Ser Ser Ser Gln Leu Phe Pro Pro Ser Arg
465                 470                 475                 480
Leu Arg Tyr Asn Arg Ser Asp Ile Met Pro Ser Gly Arg Ser Arg Leu
                485                 490                 495
Leu Glu Asp Phe Arg Asn Asn Arg Phe Pro Asn Leu Gln Leu Arg Asp
            500                 505                 510
Leu Ile Gly His Ile Val Glu Phe Ser Gln Asp Gln His Gly Ser Arg
        515                 520                 525
Phe Ile Gln Gln Lys Leu Glu Arg Ala Thr Pro Ala Glu Arg Gln Met
    530                 535                 540
Val Phe Asn Glu Ile Leu Gln Ala Ala Tyr Gln Leu Met Thr Asp Val
545                 550                 555                 560
Phe Gly Asn Tyr Val Ile Gln Lys Phe Phe Glu Phe Gly Ser Leu Asp
                565                 570                 575
Gln Lys Leu Ala Leu Ala Thr Arg Ile Arg Gly His Val Leu Pro Leu
            580                 585                 590
Ala Leu Gln Met Tyr Gly Cys Arg Val Ile Gln Lys Ala Leu Glu Ser
        595                 600                 605
Ile Ser Ser Asp Gln Gln Ser Glu Met Val Lys Glu Leu Asp Gly His
    610                 615                 620
Val Leu Lys Cys Val Lys Asp Gln Asn Gly Asn His Val Val Gln Lys
625                 630                 635                 640
Cys Ile Glu Cys Val Gln Pro Gln Ser Leu Gln Phe Ile Ile Asp Ala
                645                 650                 655
Phe Lys Gly Gln Val Phe Val Leu Ser Thr His Pro Tyr Gly Cys Arg
            660                 665                 670
Val Ile Gln Arg Ile Leu Glu His Cys Thr Ala Glu Gln Thr Leu Pro
        675                 680                 685
Ile Leu Glu Glu Leu His Gln His Thr Glu Gln Leu Val Gln Asp Gln
    690                 695                 700
Tyr Gly Asn Tyr Val Ile Gln His Val Leu Glu His Gly Arg Pro Glu
705                 710                 715                 720
Asp Lys Ser Lys Ile Val Ser Glu Ile Arg Gly Lys Val Leu Ala Leu
                725                 730                 735
Ser Gln His Lys Phe Ala Ser Asn Val Val Glu Lys Cys Val Thr His
            740                 745                 750
Ala Ser Arg Ala Glu Arg Ala Leu Leu Ile Asp Glu Val Cys Cys Gln
        755                 760                 765
Asn Asp Gly Pro His Ser Ala Leu Tyr Thr Met Met Lys Asp Gln Tyr
    770                 775                 780
Ala Asn Tyr Val Val Gln Lys Met Ile Asp Met Ala Glu Pro Ala Gln
785                 790                 795                 800
Arg Lys Ile Ile Met His Lys Ile Arg Pro His Ile Thr Thr Leu Arg
                805                 810                 815
Lys Tyr Thr Tyr Gly Lys His Ile Leu Ala Lys Leu Glu Lys Tyr Tyr
            820                 825                 830
Leu Lys Asn Ser Pro Asp Leu Gly Pro Ile Gly Gly Pro Pro Asn Gly
        835                 840                 845
Met Leu
    850
 
           
             3 
             50 
             DNA 
             Artificial Sequence 
             
               other nucleic acid from homo sapiens,
      synthesized DNA 
             
           
            3
ctctagaggc ggccgctttt tttttttttt tttttttttt tttttttttt                50
 
           
             4 
             17 
             DNA 
             Artificial Sequence 
             
               other nucleic acid from homo sapiens,
      synthesized DNA 
             
           
            4
gttttttttt ttttttc                                                    17
 
           
             5 
             10 
             DNA 
             Artificial Sequence 
             
               other nucleic acid from homo sapiens,
      synthesized DNA 
             
           
            5
agggcgtaag                                                            10
 
           
             6 
             22 
             DNA 
             Artificial Sequence 
             
               other nucleic acid from homo sapiens,
      synthesized DNA 
             
           
            6
gcatgcctat tggacttggg ga                                              22
 
           
             7 
             24 
             DNA 
             Artificial Sequence 
             
               other nucleic acid from homo sapiens,
      synthesized DNA 
             
           
            7
gttggcagtt ctgcaagtag tagt                                            24
 
           
             8 
             22 
             DNA 
             Artificial Sequence 
             
               other nucelic acid from homo sapiens,
      synthesized DNA 
             
           
            8
cccatcacca tcttccagga gc                                              22
 
           
             9 
             26 
             DNA 
             Artificial Sequence 
             
               other nucleic acid from homo sapiens,
      synthesized DNA 
             
           
            9
ttcaccacct tcttgatgtc atcata                                          26