KR100858252B1 - Kidney-specific gene HMM12 associated with development of kidney cyst - Google Patents

Abstract

The present invention is involved in the development of renal cysts (cyst), HMM12 protein specifically expressed in kidney tissue, the gene encoding the protein, a recombinant vector comprising the gene, transformants transformed with the recombinant vector The present invention relates to markers and microarrays for diagnosing the development of kidney cysts comprising the genes, antibodies to the protein and microarrays comprising the same, and methods of treating mammalian diseases using the genes.

Kidney Cyst, HMM12 Protein, Marker, Microarray

Description

Kidney-specific gene HMM12 associated with development of kidney cyst}

1 is a schematic of a system for analyzing genes for the development of cysts.

Figure 2 shows the RT-PCR results of the HMM12 gene.

Figure 3 shows protein interaction information of the HMM12 gene prepared using STRING.

The present invention is involved in the development of mammalian diseases, in particular renal cyst (cyst), HMM12 protein specifically expressed in renal tissue, the gene encoding the protein, the recombinant vector containing the gene, transformed with the recombinant vector Transformants, markers and microarrays for diagnosing the development of renal cysts comprising the gene, antibodies to the protein and microarrays comprising the same, and methods for treating mammalian kidney cysts using the genes will be.

Cyst (cyst) is a pocket-shaped space due to abnormal proliferation of epithelial cells, the body fluid fills up, the cyst spread throughout the organs and loses function, causing a number of serious complications such as end stage renal failure and cardiovascular disease. According to Boletta, A. et al., Mol Cell 6 (5): 1267-73 (2000), the formation of cysts in the kidneys is caused by PKD (Polycystic Kidney Disease). A very common hereditary disease that occurs equally regardless of gender, geographic location, or socioeconomic environment. Cysts form in the proximal and collecting ducts in the nephron of the kidneys, causing abnormal kidney function and end-stage renal failure. It leads.

PKD is a hereditary disorder in which cysts develop on both sides of the kidney, also called polycystic kidney disease or polycystic kidney disease. There are two types, pediatric and adult, which are due to autosomal recessive inheritance, and adult type due to autosomal dominant inheritance. Adult type occurs at a high frequency of about 1 person per 200 to 1,000 people. In addition, about 50% of patients progress to chronic renal failure, with an average death age of about 50 years. The pediatric type is known to have 86% of the patients diagnosed in newborns over 3 months of age, about 50% survive until 10 years old, and 80% of patients diagnosed after 1 year old have a survival rate of 15%. have. There is no specific treatment for both types, and only symptomatic treatment for various symptoms or complications is possible.

PKD's cystogenesis mechanism has been actively studied for the discovery of therapeutic agents. However, the mechanism of cyst formation in PKD has not been fully understood. To date, the cyst formation mechanism has been revealed by the same PKD gene. It is difficult to explain both the two diseases that occur: Autosomal Dominant Polycystic Kidney Disease (ADPKD) and Autosomal Recessive Polycystic Kidney Disease (ARPKD).

Korean Laid-Open Patent Publication No. 2005-0061501 describes a composition, a pharmaceutical preparation, and a method for inhibiting CFTR, which are useful for the study and treatment of cyst fibrosis transmembrane conductance regulator protein (CFTR) -mediated diseases and conditions. There is no description about.

Accordingly, the present inventors have studied the mechanism of cyst formation, revealed that the HMM12 gene, which is involved in the development of cysts, is specifically expressed in renal tissue, thereby completing the present invention.

Accordingly, it is an object of the present invention to provide an HMM12 protein that is specifically expressed in kidney tissue.

Another object of the present invention is to provide a gene encoding the protein.

Another object of the present invention is to provide a recombinant vector comprising the gene.

Another object of the present invention is to provide a transformant transformed with the vector.

Another object of the present invention is to provide a marker and a microarray comprising the gene.

Another object of the present invention is to provide an antibody against the protein and a microarray comprising the antibody.

Another object of the present invention to provide a method for treating a mammalian kidney cyst using the gene.

In order to achieve the above object, the present invention provides an HMM12 protein involved in the development of renal cysts (cysts), consisting of the amino acid sequence represented by SEQ ID NO: 2, and specifically expressed in renal tissue.

Even if a candidate group of genes involved in the cyst formation mechanism is extracted by a method such as a DNA chip, it is actually a gene involved in the cyst formation mechanism, a housekeeping gene, or an experimental error until the candidate group is knocked out and examined. It is indistinguishable. However, if the candidate group is identified as a tissue-specific gene by performing an Audic-Claverie Test based on the EST (Expressed Sequence Tag) or using an experiment such as RT-PCR, the candidate may be judged as a genetic influence factor. Reliability is increased. The inventors have provided higher reliability for candidate groups of genes involved in cyst development by examining candidate groups of genes involved in cyst development and examining whether they contain tissue specific information.

Cysts are diseases in which empty spaces are formed in tissues due to abnormal proliferation of cells, and fluids are filled and tissues lose their function. In order to determine the cause of cysts in the kidneys, the present inventors investigated the expression status of genes by DNA chip experiments using 6-, 12-, and 18-month-old mice in which cysts developed, and performed functions using UniProt database. In addition, information on protein-protein interactions was extracted from the STRING database. The inventors selected 110 genes as candidate candidates for renal specific genes using Audic-Claverie Test for EST information of genes that were significantly expressed at each expression time point. Of these genes, the HMM12 gene was indeed kidney-specific and verified by RT-PCR experiment. 1 is a schematic of a system for analyzing genes for the development of cysts. The purpose of this system is to identify whether the hourly significant genes from DNA chip experiments at 6, 12, and 18 months of cyst development are actually tissue-specific genes and to analyze information about protein-protein interactions.

Accordingly, the present invention provides a HMM12 protein involved in the development of renal cysts and specifically expressed in renal tissue. The range of HMM12 proteins according to the present invention includes proteins having the amino acid sequence represented by SEQ ID NO: 2 and functional equivalents of the proteins. "Functional equivalent" means at least 70%, preferably at least 80%, more preferably at least 90%, even more preferably at least 70% of the amino acid sequence represented by SEQ ID NO: 2 as a result of the addition, substitution, or deletion of the amino acid Is 95% or more of sequence homology, and refers to a protein that exhibits substantially homogeneous physiological activity with the protein represented by SEQ ID NO: 2.

The HMM12 protein according to the present invention is preferably derived from, but not limited to, a mouse.

The HMM12 protein according to the present invention can be used as a target of therapeutic drugs for renal cysts. When designing a drug as a treatment for cyst symptoms, HMM12 protein may reduce drug targets, allowing for faster treatment at the onset of cysts.

The present invention also provides a gene encoding the HMM12 protein. Genes of the invention include both genomic DNA and cDNA encoding the HMM12 protein. Preferably, the gene of the present invention may include the nucleotide sequence represented by SEQ ID NO: 1.

In addition, variants of the above nucleotide sequences are included within the scope of the present invention. Specifically, the gene comprises a nucleotide sequence having at least 70%, more preferably at least 80%, even more preferably at least 90%, most preferably at least 95% homology with the nucleotide sequence of SEQ ID NO: 1 can do. The "% sequence homology" for a polynucleotide is identified by comparing two optimally arranged sequences with a comparison region, wherein part of the polynucleotide sequence in the comparison region is the reference sequence (addition or deletion) for the optimal alignment of the two sequences. It may include the addition or deletion (ie, gap) compared to).

The present invention also provides a recombinant vector comprising the gene according to the present invention. The recombinant vector is preferably a recombinant animal expression vector.

The term “recombinant” refers to a cell in which a cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a protein encoded by a peptide, a heterologous peptide, or a heterologous nucleic acid. Recombinant cells can express genes or gene fragments that are not found in their natural form in either the sense or antisense form. Recombinant cells can also express genes found in natural cells, but the genes have been modified and reintroduced into cells by artificial means.

The term “vector” is used to refer to a DNA fragment (s), a nucleic acid molecule, that is delivered into a cell. Vectors can replicate DNA and be reproduced independently in host cells. The term "carrier" is often used interchangeably with "vector". The term “expression vector” refers to a recombinant DNA molecule comprising a coding sequence of interest and a suitable nucleic acid sequence necessary to express a coding sequence operably linked in a particular host organism. Promoters, enhancers, termination signals and polyadenylation signals available in eukaryotic cells are known.

In the present invention, any promoter known in the art may be used as a promoter of the expression vector. For example, the immediate early promoter of cytomegalovirus, the promoter of SV40 (SV 40 late promoter and SV 40 early promoter), the tk promoter of herpes simplex virus (HSV), and the Adeno 2 major late promoter PAdml ), Adenovirus 2 early promoter (PAdE2), p19 promoter of human parvo virus-associated virus (AAV) and metallothionein promoter of mouse, MT promoter and MMTV LTR promoter, etc. may be used, but are not limited thereto. It is not.

In the present invention, the transcription terminator of the expression vector may include any transcription terminator known in the art. For example, human growth hormone polyadenylation signals, bovine growth hormone polyadenylation signals and SV40 viral polyadenylation signals may be included.

The present invention also provides a transformant transformed with the recombinant vector according to the present invention. The transformant is preferably a mammal. Although preferably human, pets such as dogs or cats, livestock such as cattle, sheep, pigs, horses, and experimental animals such as rats, white papers, guinea pigs, and the like may also be included. In the present invention, a method of transforming a mammal with a recombinant vector includes a micro-injection method (Capecchi, MR, Cell, 22: 479 (1980)), calcium phosphate precipitation method (Graham, FL et al., Virology, 52: 456 ( 1973)), electroporation (Neumann, E. et al., EMBO J., 1: 841 (1982)), liposome-mediated transformation (Wong, TK et al., Gene, 10:87 (1980) ), DEAE-dextran treatment (Gopal, Mol. Cell Biol., 5: 1188-1190 (1985)), and gene bombardment (Yang et al., Proc. Natl. Acad. Sci., 87: 9568- 9572 (1990)), and the like.

In addition, the present invention is HMM12 according to the present invention Provided are markers for diagnosing renal cyst development, including genes. HMM12 of the present invention Since the gene can be used as a marker for diagnosing or diagnosing a genetic tendency of cyst induction, such information can be used to diagnose and perform treatment before the cyst occurs.

The present invention can also provide a kit for diagnosing the development of kidney cysts comprising the sense and antisense primers of the gene. By performing PCR amplification using the sense and antisense primers of the gene, the onset of cysts can be diagnosed through the generation of desired products. PCR conditions, sense and antisense primer lengths can be modified based on what is known in the art.

Kits of the invention may also comprise probes complementary to the gene. Hybridization may be performed using a probe complementary to the gene, and the cyst may be diagnosed through hybridization. Selection of suitable probes and hybridization conditions can be modified based on what is known in the art.

The present invention also provides a microarray for diagnosing the development of renal cyst, comprising the HMM12 gene according to the present invention. The microarray of the present invention can be easily prepared by a manufacturing method commonly used in the art using the marker of the present invention.

In the microarray of the present invention, "microarray" refers to a microarray in which a group of polynucleotides is immobilized to a high density on a substrate, and each of the polynucleotide groups is immobilized in a predetermined region. Such microarrays are well known in the art. Microarrays are disclosed, for example, in US Pat. Nos. 5,445,934 and 5,744,305, the contents of which are incorporated herein by reference.

The term “substrate” refers to any substrate to which an oligonucleotide probe can be coupled under conditions that retain hybridization properties and keep the background level of hybridization low. Typically, the substrate may be a microtiter plate, membrane (eg, nylon or nitrocellulose) or microspheres (beads) or chips. Prior to application or immobilization on the membrane, nucleic acid probes can be modified to promote immobilization or to improve hybridization efficiency. Such modifications may include homopolymer tailing, coupling with different reactive functional groups such as aliphatic groups, NH ₂ groups, SH groups and carboxyl groups, or coupling with biotin, hapten or protein.

The invention also provides an antibody against the HMM12 protein according to the invention. Antibodies of the present invention can be produced by a conventional method from the obtained protein by cloning each gene into an expression vector according to a conventional method to obtain a protein encoded by the HMM12 gene. This includes partial peptides that can be made from the protein, and the partial peptide of the present invention includes at least 7 amino acids, preferably 9 amino acids, more preferably 12 or more amino acids. The form of the antibody of the present invention is not particularly limited and a part thereof is included in the antibody of the present invention and all immunoglobulin antibodies are included as long as they are polyclonal antibody, monoclonal antibody or antigen-binding. Furthermore, the antibody of this invention also contains special antibodies, such as a humanized antibody.

The invention also provides a microarray for diagnosing the development of kidney cysts, comprising an antibody of the HMM12 protein according to the invention. The microarray of the present invention may refer to the aforementioned microarray except that the antibody of the present invention is immobilized in place of the polynucleotide in the microarray.

The invention also provides a method of treating a mammalian kidney cyst by inhibiting the expression of the HMM12 gene according to the invention in a mammal. Preferably, the mammal is a human, but may also include non-human animals such as pets such as dogs and cats, livestock such as cattle, sheep, pigs, horses, and experimental animals such as rats, white papers, guinea pigs, and the like.

As a method of inhibiting the expression of the HMM12 gene in a mammal, various methods known in the art may be used. Said "inhibition of gene expression" includes inhibition of gene transcription and translation into protein. Also included are not only complete stops of gene expression, but reduced expression.

Antisense molecules are most commonly used to inhibit the expression of certain endogenous genes in mammals. Antisense molecules inhibit the expression of target genes by inhibiting transcriptional initiation by triple chain formation, transcriptional inhibition by hybridization at sites where local open loop structure is formed by RNA polymerase, and in RNA where synthesis is in progress. Inhibition of transcription by hybrid formation, inhibition of splicing by hybridization at the junction of introns and exons, inhibition of splicing by hybridization at the site of splicosomal formation, transition from nucleus to cytoplasm by hybridization with mRNA Inhibition of translation initiation by hybridization at the site of translation initiation factor binding. They inhibit the expression of target genes by inhibiting transcription, splicing or translation processes.

The antisense molecule used in the present invention may inhibit the expression of the target gene by any of the above actions. Representative antisense molecules include triplets, ribozymes, RNAi, or antisense nucleic acids. The triple agent allows the initiation of transcription to be suppressed by winding around double helix DNA to form a three-stranded helix (Maher et al., Antisense Res. And Dev., 1 (3): 227, 1991; Helene, C., Anticancer Drug Design, 6 (6): 569, 1991). Ribozymes are RNA molecules that possess the ability to specifically cleave single stranded RNA. Ribozymes can be engineered to recognize and cleave site specific nucleotide sequences in RNA molecules (Cech, J. Amer. Med. Assn., 260: 3030, 1998). Thus, the main advantage of this method is that only mRNAs with specific sequences are inactivated because they are sequence specific. RNAi is a method of inhibiting gene expression at the transcriptional level or post-transcriptional level by using small-molecule RNA in the form of a hairpin that is specifically sequenced (Mette et al., EMBO J., 19: 5194-5201, 2000). . Antisense nucleic acid refers to a DNA or RNA molecule that is at least partially complementary to a particular mRNA molecule (Weintraub, Scientific American, 262: 40, 1990). In cells, antisense nucleic acids hybridize with the corresponding mRNAs to form double stranded molecules. This inhibits the translation of mRNA (Marcus-Sakura, Anal. Biochem., 172: 289, 1988).

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Example

Example  One: DNA  Chip experiment

According to Zoja, C. Cytogenet Genome Res 105 (2-4): 479-8 (2004), in 18-month-old mice, cysts develop in the kidneys of 6-month-old mice, and their cysts increase over time. The cysts were very large and several water clots were observed. DNA chip experiments were performed using CodeLink Bioarray: UniSet Mouse 20K I Bioarray (GE Healthcare; hppt: //www.gehealthcare.com), and Quintet (Choe, JK LNCS 3642: 392). (2005)) was used to extract a list of significant genes in the 6, 12, and 18 months following cyst changes.

Table 1 below shows the number of significant genes over time as a result of DNA chip experiments.

time Significant gene (TG / WT) Kidney Specific Related Significant Genes 6 months 1521 (845/676) 67 12 months 1220 (639/581) 51 18 months 1533 (720/813) 45

In Table 1, TG is a mouse with a cyst, and WT represents a mouse without a cyst.

Example  2: TISA  Set Up Renal Specific Gene Candidates Using Database

Noh, S.J. DNA Res 13 (5): 229-43 (2006) published a Tissue-specific Alternative Splicing (TISA) database that contains information on genes, transcript structure, alternative splicing events and tissue specificity. In order to check whether there are significant genes extracted from the TISA experiments in the TISA, check whether the Refseq information of the TISA genes is the same as the Refseq corresponding to the probe on the DNA chip, and connect the TISA database with the DNA chip. If any of the kidney specific genes predicted in the gene was significantly expressed on the DNA chip, it was extracted as a candidate group of the kidney specific genes.

Example  3: renal specific gene candidate group RT - PCR  Experiment

Mice were male and female of 9-10 weeks old B6 strain. There are 20 types of tissues that can be extracted: virgin breast, breast during lactation, breast after breast feeding, colon, ear, pancreas, kidney, liver, lung, cerebral, cerebellum, ovary, uterus, testes, thymus, stomach, spleen, Skin, heart and duodenum. Taking into account the characteristics of each tissue, the lipid, breast, cerebral, and cerebellum are crushed about 100 mg with TissueLyser (Qiagen, Valencia, CA, USA), and using the RNeasy lipid tissue mini kit (Qiagen, Valencia, CA, USA). RNA was extracted. The remaining tissue was ground to about 30 mg with the same homogenizer and RNA was obtained with the RNeasy mini kit (Qiagen, Valencia, CA, USA). From the extracted RNA 1ug, RT-PCR was performed using Standard Two-step RT-PCR- & GO ™ (Q-biogene, Cat # RT2ST100) to obtain about 100ul of cDNA. Primers for these genes to identify kidney specific genes (forward primer: 5'-TGAGCCCACTGATAAGCATGA-3 '(SEQ ID NO: 3) and reverse primers: 5'-TTTGCAATGTCCTGGAGGTG-3' (SEQ ID NO: 4)) (PCR product size 536 bp) and PCR was performed using cDNA for the 20 tissues obtained above. PCR conditions were last extended for 30 min of predenatured 95 ° C. 5 min, followed by 30 cycles of 95 ° C. 1 min, annealing 58 ° C. 1 min, extension 72 ° C. 1 min, and 72 ° C. 5 min. 1% agarose gel electrophoresis was performed to confirm the PCR product.

Figure 2 shows the RT-PCR results of the HMM12 gene. Description of each lane of Figure 2 is as follows:

Br1: virgin breast Ce2: cerebral Th: thymus Br2: breast during lactation Pa: Pancreas Ov: Ovary Br3: breast after lactation Ki: height Li: liver Co: Colon Sk: skin Ut: Uterus Ea: Ear He: heart Sp: Spleen Du: duodenum Te: Testicle St: above Ce1: Cerebellum Lu: lung

As can be seen in Figure 2, it was found that the HMM12 gene is rarely expressed in other tissues, but specifically expressed in the kidney.

Example  4: UniProt Reference of Kidney Specific Gene Candidates

UniProt (http://www.ebi.uniprot.org) was developed in 2002 as an EBI / SIB Swiss-Prot, TrEMBL database PIR-Protein Sequence Database (PIR-PSD). It is a way to combine the contents into a UniProt consortium and provide them with mutual cooperation. The consortium provides comprehensive, accurate information about protein sequences, and maintains an accessible interface despite providing many references.

To identify which protein candidates are specific to kidneys and which functions are linked to UniProt, the sequence data of kidney-specific gene candidates was applied to BLAST alignment provided by UniProt's website and identified for accuracy. Only data with values greater than 95% were adopted.

Example  5: STRING Analysis of Protein Interaction in Renal Specific Gene Candidates

STRING (Search Tool for the Retrieval of Interacting Proteins; http://string.embl.de) is a database that provides comprehensive information on protein interactions.In 2000, Snel B 2000 Nucleic Acids Res 28 (18) 3442 Version 7 was released in 2007 as a continual upgrade since its release by -4.

In order to confirm the protein interaction information of the kidney-specific gene candidate group, the mouse sequence in STRING was determined by using the candidate data and BLAST alignment (identity> = 84% and coverage> = 60%), Corresponding protein interaction information was extracted.

Figure 3 shows protein interaction information of the HMM12 gene prepared using STRING. In FIG. 3, red dots represent proteins aligned with 100% identity and 100% coverage on HMM12 and BLAST alignment. Proteins that interact with the HMM12 protein are proteins that act as transporters of amino acids, which is an important function of the kidney.

To determine the cause of the cyst's development, it is important to identify information about the protein-protein interactions, pathways, or regulatory networks involved in the cyst's development. However, analyzing and generating individual genes to reveal this information takes a very long time to identify the genes actually involved in the development of cysts.

The present inventors provide the fact that the HMM12 gene involved in the development of cysts is specifically expressed in renal tissue, allowing researchers to easily access information such as protein-protein networks or pathways for the development of cysts. The fact that it is a kidney-specific gene can reduce false positives in studies of cyst symptoms. In addition, the HMM12 gene can be used as a marker for the diagnosis of cysts, making it easier to diagnose, and when designing a drug as a therapeutic agent for cyst symptoms, the drug target for HMM12 can be reduced. Faster treatment is possible early in the onset of cysts.

<110> Korea Research Institute of Bioscience and Biotechnology <120> Kidney-specific gene HMM12 associated with development of kidney          cyst <160> 4 <170> KopatentIn 1.71 <210> 1 <211> 4102 <212> DNA <213> Mouse C57 BL / 6 <400> 1 acacaaggaa tactcacagg gttaatcctt ctgacaccaa gtcacacttt aactcattgt 60 catcaggaca aagattaaac gctgcctgcg taagagtcag ggctccagca gaccctgaaa 120 gctgagctgt ccagaccccc gaagtgaaga aaagaggcga gggcaaggga gggccagaac 180 caggggagag agaaaggagg ggcagcccac cagcccgctg tctcgccaca gaaccggctc 240 agctccaact ccaggagtca ctcagctgca gaggcagtgg cagccctact cctcaggcaa 300 agggcagcag acagacagac agaggtccta ggactggagg tcctcaggca ttgaccactc 360 agcctggccc agccccatgg ccttcaatga cctcctgaaa caggtggggg gtgtcggacg 420 cttccagctg atccaggtca ccatggtggt tgctccccta ctgctgatgg cttcccacaa 480 caccctgcag aacttcactg ctgctatccc cgctcaccac tgccgcccac ctgctaatgc 540 caacctcagc aaagatggag gtctggaggc ctggctgccc ctggacaagc aaggacgacc 600 cgaatcttgc ctccgctttc ctttcccgca caatggcaca gaggccaatg gcaccggagt 660 cacagagccc tgccttgatg gctgggtcta tgacaacagc accttccctt caaccatcgt 720 gactgagtgg aaccttgtgt gctctcatcg ggccttccgc cagctggccc agtccctgtt 780 catggtggga gtgttactgg gagccatgat gtttggctac ctggcggaca ggctgggccg 840 tcggaaggtg ctgatcttga actacctgca aacagctgtg tcgggaacct gtgcagccta 900 tgcacccaac tatactgtct actgcatttt ccggctcctc tcgggcatgt ctttggctag 960 cattgcaatc aactgcatga cactaaacat ggagtggatg cctattcaca cccgtgccta 1020 tgtgggcacc ttgattggct atgtctacag cttgggccag ttcctcctgg ctggtatcgc 1080 ctatgctgtg ccccactggc gccacctgca gcttgcggtc tctgtgcctt ttttcgttgc 1140 cttcatctac tcttggttct tcattgagtc agcccgctgg tactcctcct ctggaaggct 1200 ggacctcacc ctccgagccc tgcagagagt ggcccgcatc aacgggaaac aagaagaggg 1260 ggctaagctg agtatagagg tgctccagac cagcctgcag aaggaactga ctctaaacaa 1320 aggccaggcc tccgccatgg agctgctgcg ctgccccacg cttcggcgcc tcttcctctg 1380 cctctctatg ctgtggtttg ccactagctt tgcctactac gggctggtca tggacctgca 1440 gggctttggg gtcagcatgt accttatcca ggtgattttc ggcgctgtgg acctgcctgc 1500 caagtttgtg tgcttcctag tcatcaattc catgggccgc cggcctgcac agttggcctc 1560 cctgctgctg gcaggcatct gtatcctagt gaatggcata ataccgaggg gccatacaat 1620 cattcgcaca tccctggctg tactagggaa aggctgtctg gcttcctctt tcaactgcat 1680 cttcctgtac accggagagc tgtaccccac aatgattcgg cagacgggcc tgggcatggg 1740 cagcaccatg gcccgggtgg gcagcatagt gagcccactg ataagcatga ctgccgagtt 1800 ctacccctcc atacctctct tcatcttcgg cgctgtcccc gtggccgcca gcgctgtcac 1860 tgccctgctg ccagagacct tgggccagcc gctgcctgat acagtgcagg acctgaagag 1920 caggagcaga ggaaagcaga agcaacagca gctggaacag cagaagcaga tgataccact 1980 ccaggtctca acacaagaga agaacggact ctgaaaatgg agagggcgtc acacagcact 2040 aaagggagtg gggttctgca ggtcccacca tctatgagga gggggagtga gtggagggac 2100 tgggccatct aaatgcggag cctgccattc agagaaatgc ctccccaaag gtcacttcaa 2160 tagacctact aggaacaaaa gctctaactg tgtgcagctt cttaagcaga atgagctctt 2220 cctcagccat cttccagctc agtggtcact cctccagctc ctgtcccttc cacctccagg 2280 acattgcaaa gaatcccaga caattaaata aatttcttct accttggtcc ctctctcttc 2340 ttctctacct ctcctatcca accatttctg ttttaactac atgttttctg tctcttccct 2400 ttgcagaaat cagaatttaa tgagaaactg aagtgaaatc agcagtcaga ggctctgccc 2460 tgggctcttc tcctgctgta agagcagaca gaaacttcta gcagcaaaag agatagatgc 2520 ttataggcgt gatgtcagga tcctgtcccc tgcaagttca gaccactagc caacctaatc 2580 tttcctgtgt ttgatgggtt ttcaacgttg ctctgcttcc tgtaggttca ggggctggtg 2640 gaggagtgag acgaggacca gacagtattt aacagccagg gagaccaaac actgagtgtt 2700 aattatatgc caggtggtag cttcagcaat ttaaatgtat taattcagtt tatcctcttt 2760 ggtttcattt tcttagtttt cctgataagg ggtcttatct agctagttat tttgttaact 2820 tgacacaggc tagagtcatt tgaaaggaaa gaacctcaac tgaaaaaaaa tgtctctata 2880 agtggcctga aggcaagcct gtgcagggca gattcttaat taatgatcga tgtaggagga 2940 ttcagaccac tgttgacaac actgcccctg ggatgatggt cctgtttgct ataacaaagc 3000 aagctgagca agccagtgag cagtgttctt ccacagcctc tgctacagtc tctgccccga 3060 cttcccacca tgtcagagtg tgacctgaga gctgtaagat gaaacaaacc ctttcctctc 3120 taagtagctt ttggtcatgg tattttgtca cagcaatgga aatactaaga cagggtctca 3180 tccctagctg gccttgacct gactaagtag ctgagggtga ctttgaacct tgagccttct 3240 gcctccacct cccaaatgca agcattccag gcgtgtgccg caatgctcaa atctgaaaat 3300 agtacccttt gattcctatc ttggtctctt ttcagtcagt gtgtcaggga ccttgtagca 3360 tcttctgtga cttgaccaag atggaacact ctgaattctc aacttcctgg tcctggtaga 3420 gttctcctcc tgctcagggc ttcacctcat cattccaggt tccacacagg cttttagtcc 3480 aaagcccttc cttacaggat ttttcctcct tcaaagatgt ttcccgaccc cagagagatt 3540 tagaggacag agtctctacc agctaagctt cctgccctag atcaggctgt gcagaggaac 3600 cctagacgag gtctacacaa actccttgga actttcaatt gtgtggccta aaacataaat 3660 gacatgcaga tataattaaa ttaatgattt cgaagtgaag aaactatctt aaattatttg 3720 ggttagccca atatgattac aaaggcagtt agacagatca gtggacagca gatataacag 3780 aagaatcaag agggtggagt gactcaagaa ggaatgaaaa ataaaaatca gttcaagaca 3840 gagacaataa gcttcaatta gttgttagca aattaaagta acttctgatc attttcaatc 3900 aaatagatac atttagcatt atcaaatgaa taaatatgtg gtatgggcct ccaggccaag 3960 gaatgaaact tccgggagct taaataggaa ggaaatggat tcttctttga aaccaccaaa 4020 aggaacacat ccctgttgac atagcaacct tcaatatcca gcctctaact atacaagaat 4080 aaactggact gttttaagcc tc 4102 <210> 2 <211> 545 <212> PRT <213> Mouse C57 BL / 6 <400> 2 Met Ala Phe Asn Asp Leu Leu Lys Gln Val Gly Gly Val Gly Arg Phe   1 5 10 15 Gln Leu Ile Gln Val Thr Met Val Val Ala Pro Leu Leu Leu Met Ala              20 25 30 Ser His Asn Thr Leu Gln Asn Phe Thr Ala Ala Ile Pro Ala His His          35 40 45 Cys Arg Pro Pro Ala Asn Ala Asn Leu Ser Lys Asp Gly Gly Leu Glu      50 55 60 Ala Trp Leu Pro Leu Asp Lys Gln Gly Arg Pro Glu Ser Cys Leu Arg  65 70 75 80 Phe Pro Phe Pro His Asn Gly Thr Glu Ala Asn Gly Thr Gly Val Thr                  85 90 95 Glu Pro Cys Leu Asp Gly Trp Val Tyr Asp Asn Ser Thr Phe Pro Ser             100 105 110 Thr Ile Val Thr Glu Trp Asn Leu Val Cys Ser His Arg Ala Phe Arg         115 120 125 Gln Leu Ala Gln Ser Leu Phe Met Val Gly Val Leu Leu Gly Ala Met     130 135 140 Met Phe Gly Tyr Leu Ala Asp Arg Leu Gly Arg Arg Lys Val Leu Ile 145 150 155 160 Leu Asn Tyr Leu Gln Thr Ala Val Ser Gly Thr Cys Ala Ala Tyr Ala                 165 170 175 Pro Asn Tyr Thr Val Tyr Cys Ile Phe Arg Leu Leu Ser Gly Met Ser             180 185 190 Leu Ala Ser Ile Ala Ile Asn Cys Met Thr Leu Asn Met Glu Trp Met         195 200 205 Pro Ile His Thr Arg Ala Tyr Val Gly Thr Leu Ile Gly Tyr Val Tyr     210 215 220 Ser Leu Gly Gln Phe Leu Leu Ala Gly Ile Ala Tyr Ala Val Pro His 225 230 235 240 Trp Arg His Leu Gln Leu Ala Val Ser Val Pro Phe Phe Val Ala Phe                 245 250 255 Ile Tyr Ser Trp Phe Phe Ile Glu Ser Ala Arg Trp Tyr Ser Ser Ser             260 265 270 Gly Arg Leu Asp Leu Thr Leu Arg Ala Leu Gln Arg Val Ala Arg Ile         275 280 285 Asn Gly Lys Gln Glu Glu Gly Ala Lys Leu Ser Ile Glu Val Leu Gln     290 295 300 Thr Ser Leu Gln Lys Glu Leu Thr Leu Asn Lys Gly Gln Ala Ser Ala 305 310 315 320 Met Glu Leu Leu Arg Cys Pro Thr Leu Arg Arg Leu Phe Leu Cys Leu                 325 330 335 Ser Met Leu Trp Phe Ala Thr Ser Phe Ala Tyr Tyr Gly Leu Val Met             340 345 350 Asp Leu Gln Gly Phe Gly Val Ser Met Tyr Leu Ile Gln Val Ile Phe         355 360 365 Gly Ala Val Asp Leu Pro Ala Lys Phe Val Cys Phe Leu Val Ile Asn     370 375 380 Ser Met Gly Arg Arg Pro Ala Gln Leu Ala Ser Leu Leu Leu Ala Gly 385 390 395 400 Ile Cys Ile Leu Val Asn Gly Ile Ile Pro Arg Gly His Thr Ile Ile                 405 410 415 Arg Thr Ser Leu Ala Val Leu Gly Lys Gly Cys Leu Ala Ser Ser Phe             420 425 430 Asn Cys Ile Phe Leu Tyr Thr Gly Glu Leu Tyr Pro Thr Met Ile Arg         435 440 445 Gln Thr Gly Leu Gly Met Gly Ser Thr Met Ala Arg Val Gly Ser Ile     450 455 460 Val Ser Pro Leu Ile Ser Met Thr Ala Glu Phe Tyr Pro Ser Ile Pro 465 470 475 480 Leu Phe Ile Phe Gly Ala Val Pro Val Ala Ala Ser Ala Val Thr Ala                 485 490 495 Leu Leu Pro Glu Thr Leu Gly Gln Pro Leu Pro Asp Thr Val Gln Asp             500 505 510 Leu Lys Ser Arg Ser Arg Gly Lys Gln Lys Gln Gln Gln Leu Glu Gln         515 520 525 Gln Lys Gln Met Ile Pro Leu Gln Val Ser Thr Gln Glu Lys Asn Gly     530 535 540 Leu 545 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 3 tgagcccact gataagcatg a 21 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 tttgcaatgt cctggaggtg 20  

Claims (13)

delete delete delete delete delete delete delete delete Microarray for diagnosing the development of kidney cyst, comprising the nucleotide sequence represented by SEQ ID NO: 1. delete A microarray for diagnosing the development of renal cyst, comprising an antibody to HMM12 protein involved in the development of renal cyst (cyst), consisting of the amino acid sequence represented by SEQ ID NO: 2, and which is superior in renal tissue only. delete delete

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