KR100794706B1 - Probes or primers for diagnosing obesity or diabetes derived from genes coding deubiquitinating enzymes and methods for diagnosing obesity or diabetes using the same - Google Patents

Abstract

Probes or primers derived from genes encoding deubiquitinating enzymes are provided to identify molecules mediating decomposition of materials which are associated with the insulin signal transfer, so that they are useful for diagnosing diseases associated with insulin signal transfer such as obesity or diabetes. A probe for diagnosing obesity or diabetes comprises 10-100 consecutive nucleotides or their complementary nucleotides selected from genes of SEQ ID NO:1 to SEQ ID NO:16 encoding deubiquitinating enzymes. A microarray comprises the probe for diagnosing obesity or diabetes. A method for diagnosing obesity or diabetes comprises the steps of: collecting a nucleic acid sample from adipocytes of normal person and a testing person; measuring the expression amount of deubiquitinating enzyme genes in the sample; and deciding the obesity or diabetes when the expression amount of deubiquitinating enzyme genes for the testing person is smaller than that of normal person.

Description

Probes or primers for diagnosing obesity or diabetes derived from genes coding deubiquitinating enzymes and methods for diagnosing obesity or diabetic diabetic or diabetic diagnostic probes, primers, or methods for diagnosing obesity or diabetes using the same using the same}

Figure 1 shows the results of measuring the expression level of deubiquitase in the insulin-treated adipocytes and untreated adipocytes by reverse transcriptase polymerase chain reaction.

Figure 2 shows the result of measuring the relative expression of deubiquitination enzyme in insulin-treated adipocytes and untreated adipocytes by real-time polymerase chain reaction.

The present invention relates to a probe and primer for diagnosing obesity or diabetes, and also to a method for diagnosing obesity or diabetes using the same.

Insulin is a hormone secreted by the pancreas that regulates energy functions such as sugar and fat metabolism. Tyrosine groups in the insulin receptor are phosphorylated when insulin binds to insulin receptors in insulin-sensitive tissues such as muscle, fat cells, and liver. This phosphorylated insulin receptor induces phosphorylation of target proteins such as the insulin receptor substrate (IRS) in the cell. Insulin receptor substrates induce phosphatidyl inositol 3 kinase, which phosphorylates PtdIns (3,4) P2 to produce PtdIns (3,4,5) P3. Phosphatidyl kinase is a major signaling pathway in insulin signaling. Another major signaling process is the mitogen activated protein kinase pathway. Insulin receptor substrates also interact with protein-tyrosine phosphatases containing SH2 domains such as GRB2 and SHP2. GRB2 interacts with guanine nucleotide exchange factors mSOS, Ras, Raf, and MEK, and these substances activate the MAPK process. Insulin is highly expressed in liver, muscle and adipocytes and is known to play an important role in the differentiation of adipocytes. Fat cells play an important role in maintaining the metabolism and homeostasis of the entire body, and in particular, homeostasis also plays an important role in insulin resistance.

Fat stored in fat cells is used as an important energy source in the body. However, as obesity progresses, not only does the adipocytes increase in number, but the synthesis of a large amount of triglycerides by the differentiation of excessive adipocytes is accompanied by morphological changes including the size of the adipocytes and changes in various gene expressions. Adipocyte differentiation is promoted by stimulation of insulin, insulin like growth factor-1, growth hormone, and the like, and in the process, CCAAT enhancer-binding protein (C / EBP) family, peroxisome proliferator-activated receptor ( PPAR) gamma and other transcription factors are observed to increase. These transcription factors, along with adipocyte regulators, promote the differentiation of adipocytes and increase the expression levels of enzymes such as fatty acid binding proteins aP2 and fatty acid synthase. On the other hand, in the case of type 2 diabetes in which obesity progresses to diabetes, palmitate, a triglyceride, gradually increases insulin resistance, and ultimately destroys pancreatic beta cells to induce diabetes. Proc. Natl. Acad. Sci. USA. 95, 2498-2502 (1998). On the other hand, it is reported that excessive triglyceride accumulation is involved in the progression of fatty liver [J. Clin. Invest., 98, 1575-1584 (1996).

On the other hand, numerous metabolisms in organisms are regulated by ubiquitination and deubiquitination. The ubiquitin (Ub) is composed of 76 amino acids, a relatively small molecular weight of 8,565 Daltons present in all eukaryotic cells, and is involved in numerous cell functions such as selective degradation of proteins and regulation of cell division. (Annu. Rev. Nutr. 15, 161-189 (1995); Annu. Rev. Biochem. 61, 761-807 (1992); Annu. Rev. Genet. 26, 179-207 (1992); Proc. Natl. Acad.Sci. USA.88, 4606-4601 (1991); Nature 349, 132-138 (1991); Annu. Rev. Cell Biol. 3, 1-30 (1987); J. Mol. Cell. Biol. 6 4602-4610 (1986). Ubiquitin-mediated proteolysis begins by binding ubiquitin to target proteins. This process involves ubiquitin activating enzyme (E1), ubiquitin carrier proteins (E2), ubiquitin ligase (E3), and ubiquitin factors (E4). Ubiquitinylated proteins are recognized by the 26S proteasome and the reaction proceeds ATP dependent. These ubiquitinated proteins can be deubiquitinated to prevent protein degradation. This deubiquitination is carried out by a deubiquitination enzyme, which serves to cut ubiquitin from a protein to which ubiquitin is bound, and the ubiquitin that is cut can be combined with another protein. Deubiquitination enzymes include C-terminal hydrolases (UCH), the ubiquitin specific processing proteases (UBP or USP), Jab1 / Pad1 / MPN domain-containing metallo enzymes (JAMM), Otu-domain ubiquitin-aldehyde-binding proteins (Otubain) , Ataxin-3 / Josephin, and the like.

Recently, some of the substances involved in insulin signaling are known to be degraded by ubiquitin. Examples include IRS, peroxisome proliferator activated receptor α (PPARα), forkhead transcription factor (FOXO1), sterol regulatory element binding proteins (SREBP) (Diabetes. 48, 1359-1364 (1999); J. Biol. Chem. 277 37254-37259 (2002); Proc. Natl. Acad. Sci. USA. 100, 11285-11290 (2003); Proc. Natl. Acad. Sci. USA. 100, 13833-13838 (2003)). However, no research has been done at the molecular or genetic level as to how the degradation by ubiquitin occurs.

Therefore, there is a need in the art to identify which molecules are mediated by the degradation of substances involved in insulin signaling, and when identifying the molecule or gene encoding it, diseases associated with insulin signaling, e.g. It is expected to be useful for diagnosing obesity or diabetes.

The inventors of the present invention have been researching to find out which molecules are mediated by the decomposition of substances involved in insulin signaling. Surprisingly, the amount of gene expression of a specific deubiquitase is significantly increased when adipose progenitor cells are treated with insulin. It was found to decrease.

Accordingly, an object of the present invention is to provide a probe for diagnosing obesity or diabetes or a microarray including the same, which is derived from a gene encoding a specific deubiquitination enzyme.

Another object of the present invention is to provide an amplification primer pair for diagnosing obesity or diabetes, which is derived from a gene encoding the deubiquitination enzyme.

In addition, an object of the present invention is to provide a method for diagnosing obesity or diabetes, comprising measuring the expression level of the gene encoding the deubiquitination enzyme.

According to one aspect of the present invention, there is provided a diagnostic probe for obesity or diabetes, consisting of 10 to 100 consecutive nucleotides or complementary nucleotides selected from genes encoding deubiquitination enzymes selected from the group consisting of polynucleotides of SEQ ID NOs: 1-16 Is provided.

According to another aspect of the invention, there is provided a microarray comprising the probe for diagnosing obesity or diabetes.

According to another aspect of the present invention, diagnosing obesity or diabetes, consisting of 10 to 100 consecutive nucleotides and complementary nucleotides selected from genes encoding deubiquitination enzymes selected from the group consisting of polynucleotides of SEQ ID NOs: 1-16 Primer pairs for amplification are provided.

According to still another aspect of the present invention, there is provided a method of preparing a nucleic acid sample from adipocytes isolated from a normal person and a subject, respectively; Measuring the expression level of a gene encoding at least one deubiquitination enzyme selected from the group consisting of polynucleotides of SEQ ID NOs: 1 to 16 in each of the obtained nucleic acid samples; And obesity or diabetes when the expression amount of the gene encoding the deubiquitase enzyme in the nucleic acid sample obtained from the adipocytes of the subject is less than the expression amount of the gene encoding the deubiquitase enzyme in the nucleic acid sample obtained from the adipocytes of normal persons. There is provided a method for diagnosing obesity or diabetes, comprising the step of determining.

Hereinafter, the present invention will be described in detail.

The present inventors focused on deubiquitination enzymes while conducting studies to determine which molecules are mediated by the degradation of substances involved in insulin signaling, and specific deubiquitination was observed when adipocytes were treated with insulin. It was found that the amount of gene expression of the enzyme decreased significantly. In other words, as a result of measuring the difference in the expression of the deubiquitase enzyme gene by insulin treatment in adipose progenitor cells, it was surprisingly found that the expression level of 16 deubiquitase enzyme genes among 55 deubiquitase enzyme genes significantly decreased. . Therefore, when the expression level of the 16 deubiquitination enzymes is measured, obesity or diabetes can be diagnosed.

Deubiquitination enzymes exhibiting a significantly decreased expression level when treated with adipose progenitor cells are shown in Table 1 below.

SEQ ID NO: Gene name NCBI GenBank Registration Number One USP7 NM_001003918 2 USP10 NM_009462 3 USP11 NM_145628 4 USP16 NM_024258 5 USP18 NM_011909 6 USP20 NM_028846 7 USP22 NM_001004143 8 USP25 NM_013918 9 USP28 BC088733 10 USP36 XM_126772 11 USP38 NM_027554 12 USP39 NM_138592 13 USP46 NM_177561 14 USP47 NM_133758 15 USP53 NM_133857 16 USP54 AK047620

Accordingly, the present invention provides a probe for diagnosing obesity or diabetes, consisting of 10 to 100 consecutive nucleotides or complementary nucleotides selected from genes encoding deubiquitination enzymes selected from the group consisting of polynucleotides of SEQ ID NOs: 1-16.

The probe may be prepared using a conventional method in the art, based on the gene encoding the deubiquitination enzyme, ie, the gene sequence of SEQ ID NOs: 1 to 16, the probe sequence may have a length of 10 to 100, Preferably it is 10-50 pieces, More preferably, it may be 10-30 pieces.

In addition, the nucleotide or the complementary nucleotide is a sequence capable of detecting the gene encoding the deubiquitination enzyme, may be selected from the group consisting of SEQ ID NO: 17 to 48, wherein SEQ ID NO: 17, 18, 21, The sequences of 22, 27, 28, 35 or 36 can be preferably used as a probe for diagnosing obesity or diabetes.

The present invention includes a microarray comprising the probe for diagnosing obesity or diabetes. The microarrays can be prepared by conventional methods in the art using the probes according to the invention.

The present invention also provides amplification primer pairs for diagnosing obesity or diabetes, consisting of 10 to 100 contiguous nucleotides and complementary nucleotides selected from genes encoding deubiquitination enzymes selected from the group consisting of polynucleotides of SEQ ID NOs: 1-16. It includes.

“Primer” herein refers to the synthesis of template-directed DNA synthesis under appropriate conditions in suitable buffers (eg, four different nucleoside triphosphates (ATP, GTP, CTP, TTP) and DNA polymerase) and at appropriate temperatures. It refers to a single stranded oligonucleotide that can act as a starting point. The appropriate length of the primer may vary depending on the purpose of use, but is 10 to 100, preferably 10 to 50, more preferably 10 to 30 nucleotides. Short primer molecules generally require lower temperatures to form stable hybrids with the template. The primer sequence need not be completely complementary to the template, but should be sufficiently complementary to hybridize with the template. The primer hybridizes to a target DNA, ie, a gene encoding a deubiquitination enzyme selected from the polynucleotides of SEQ ID NOS: 1-16, and initiates amplification in an allelic form of DNA in which the primer exhibits complete homology. This primer is used in pairs with a second primer that hybridizes to the other side. The product is amplified from two primers by amplification, and only the amplified product is specifically stained and visualized. This means that certain allelic forms exist. If the allele-specific polynucleotides used herein undergo different denaturation processes, they may be used by other methods as well as Infinium ^™ Assay.

The primer pair is SEQ ID NOs: 17 and 18; SEQ ID NOs: 19 and 20; SEQ ID NOs: 21 and 22; SEQ ID NOs: 23 and 24; SEQ ID NOs: 25 and 26; SEQ ID NOs: 27 and 28; SEQ ID NOs: 29 and 30; SEQ ID NOs: 31 and 32; SEQ ID NOs: 33 and 34; SEQ ID NOs: 35 and 36; SEQ ID NOs: 37 and 38; SEQ ID NOs: 39 and 40; SEQ ID NOs: 41 and 42; SEQ ID NOs: 43 and 44; SEQ ID NOs: 45 and 46; And SEQ ID NOs: 47 and 48, more preferably SEQ ID NOs: 17 and 18; SEQ ID NOs: 21 and 22; SEQ ID NOs: 27 and 28; And SEQ ID NOs: 35 and 36.

Sequences of SEQ ID NOs: 17 to 48 that can be preferably used as the probes and primers are shown in Table 2 below.

SEQ ID NO: order 17 5'-GGGATGGCAAATGGTGTA-3 ' 18 5'-TCCTCTGCGACTATCTGC-3 ' 19 5'-TTCAAGCACACTGAACCC-3 ' 20 5'-TGGCATGGCCATTGACCA-3 ' 21 5'-AAAGATGGCACTTGGCCC-3 ' 22 5'-CCAACCTTGTTCTTGAACA-3 ' 23 5'-CATGGGAAAGAAACGGAC-3 ' 24 5'-CTCCTGAGAATTTCTGCC-3 ' 25 5'-CCTGGAAGTGAAGTCGTG-3 ' 26 5'-CAAGGAGTTAAGGCAGCA-3 ' 27 5'-CCTATTGCTGTGGCTGAT-3 ' 28 5'-GGCATAGCCTCGGAACAT-3 ' 29 5'-CAGCTTCAAGGTGGACAA-3 ' 30 5'-AGATGTAGTCCTGGCACA-3 ' 31 5'-TATCTAGAGCAGCCATCAA-3 ' 32 5'-GCCTGGTTCTGGATAAAG-3 ' 33 5'-CCCACCTCTCACAGTGAT-3 ' 34 5'-TACACAGACACTTTTCGGA-3 ' 35 5'-AAGGACTCGGCTGATGAT-3 ' 36 5'-GGGGAAAAGCACTTTCTG-3 ' 37 5'-CCTTGTGCAGCATATTCC-3 ' 38 5'-AGAACTGCAAGAGCACCA-3 ' 39 5'-CAAGTACTTTCAAGGCCG-3 ' 40 5'-TGGTACCATCATATGCCC-3 ' 41 5'-GGATGAGGGTAAAAAAGCT-3 ' 42 5'-CTTCCACAGTGAACGACC-3 ' 43 5'-TGTTGAAAGCTCCGAGAC-3 ' 44 5'-CTGCTGTTGCTGCAGTGA-3 ' 45 5'-GATATGACACAGACAGCAG-3 ' 46 5'-AAGGGAACTTCTGCTTCC-3 ' 47 5'-GGTAGTGTACAAGGGATGTT-3 ' 48 5'-GAGAGTGTCAGATGGAAGC-3 '

The present invention also comprises the steps of obtaining a nucleic acid sample from adipocytes isolated from normal and subject, respectively; Measuring the expression level of a gene encoding at least one deubiquitination enzyme selected from the group consisting of polynucleotides of SEQ ID NOs: 1 to 16 in each of the obtained nucleic acid samples; And obesity or diabetes when the expression amount of the gene encoding the deubiquitase enzyme in the nucleic acid sample obtained from the adipocytes of the subject is less than the expression amount of the gene encoding the deubiquitase enzyme in the nucleic acid sample obtained from the adipocytes of normal persons. It includes a method of diagnosing obesity or diabetes comprising the step of determining.

The method for separating fat cells from normal persons and subjects may be obtained from blood collected for various tests in a medical institution, or may be obtained from adipose tissue discarded by a fat removal procedure. For example, as disclosed in WO2000 / 53795 and WO2005 / 042730, liposuction and sedimentation from adipose tissue, enzyme treatment such as collagenase, and centrifugation Adipose cells can be separated through a process such as removing floating cells such as red blood cells by separation.

In the step of measuring the expression level of the gene encoding the deubiquitination enzyme in the nucleic acid sample, it is more preferable to measure the expression level of the gene encoding the deubiquitination enzyme of SEQ ID NO: 1, 3, 6, or 10.

In addition, the step of measuring the expression level of the gene encoding the deubiquitination enzyme is SEQ ID NO: 17 and 18; SEQ ID NOs: 19 and 20; SEQ ID NOs: 21 and 22; SEQ ID NOs: 23 and 24; SEQ ID NOs: 25 and 26; SEQ ID NOs: 27 and 28; SEQ ID NOs: 29 and 30; SEQ ID NOs: 31 and 32; SEQ ID NOs: 33 and 34; SEQ ID NOs: 35 and 36; SEQ ID NOs: 37 and 38; SEQ ID NOs: 39 and 40; SEQ ID NOs: 41 and 42; SEQ ID NOs: 43 and 44; SEQ ID NOs: 45 and 46; And a reverse transcriptase-polymerase chain reaction or a real time-polymerase chain reaction using a primer pair selected from the group consisting of SEQ ID NOs: 47 and 48.

Hereinafter, the present invention will be described in more detail with reference to Examples. However, these examples are for illustrating the present invention, and the scope of the present invention is not limited to these examples.

Example  1. Cell Culture and Differentiation into Adipocytes

Mouse adipocytes (3T3-L1 preadipocytes) were cultured in DMEM culture containing 10% fetal bovine serum (FBS), 100 U / mL penicillin, 0.1 mg / mL streptomycin sulfate. In order to differentiate from adipocytes to adipocytes, two days after the adipocytes were grown on the culture plate, in DMEM cultures containing 10% FBS, 1 μM dexamethasone, 0.5 mM methylisobutylxanthine, and 1 μM insulin After incubation for 2 days, DMEM culture medium was added every 10 days with 10% FBS, 1 μM insulin. After about 8 days after the start of differentiation, if more than 90% of the cells accumulate fat, the differentiation was considered to be over.

Example  2. Treatment of insulin with fat precursor cells and RNA  detach

Adipose progenitor cells were inoculated at 1 × 10 ⁵ cells / mL in a 100Φ culture dish and incubated for 2 days. Thereafter, the culture solution in which 100 μM of insulin was added to the culture medium was replaced, and then cultured for 12 hours. Total intracellular RNA of insulin-treated adipocytes and non-insulin-treated adipocytes was isolated using TRIzol ^™ reagent (Invitrogen, USA). The isolated RNA was dissolved in 50 μL of DEPC treated water and its concentration was measured at UV 260 nm. Initial cDNA was synthesized with 2 μg of RNA with Superscript ^™ II (Invitrogen, USA) according to the manufacturer's instructions. These cDNAs were used as template strands for reverse transcription polymerase chain reaction and real time polymerase chain reaction.

Example  3. Deubiquitination  Specific for enzyme genes primer  Making of sets

All genes of deubiquitase have a common site called UCH, regardless of length, so that the PCR product length is less than about 350 bp at the site specific to the gene of each deubiquitase except that site. Primer sets were prepared. Each gene and NCBI GenBank accession numbers are shown in Tables 3a and 3b. In addition, the primer sets produced for each gene are shown in Tables 4A to 3D.

Gene name NCBI GenBank Registration Number USP1 NM_146144 USP2 NM_016808 USP3 NM_144937 USP4 NM_011678 USP5 NM_013700 USP6 NM_004505 USP7 NM_001003918 USP8 NM_019729 USP9X NM_009481 USP9Y NM_148943 USP10 NM_009462 USP11 NM_145628 USP12 NM_011669 USP13 NM_001013024 USP14 NM_021522 USP15 NM_027604 USP16 NM_024258 USP17 NM_001033494 USP18 NM_011909 USP19 NM_027804 USP20 NM_028846 USP21 NM_013919 USP22 NM_001004143 USP24 XM_131566 USP25 NM_013918

Gene name NCBI GenBank Registration Number USP26 NM_031388 USP27 NM_019461 USP28 BC088733 USP29 NM_021323 USP30 XM_149655 USP31 XM_485998 USP32 XM_110937 USP33 NM_133247 USP34 XM_483996 USP35 XM_145945 USP36 XM_126772 USP37 NM_176972 USP38 NM_027554 USP39 NM_138592 USP40 BC089376 USP41 XM_036729 USP42 AK010801 USP43 NM_173754 USP44 NM_183199 USP45 NM_152825 USP46 NM_177561 USP47 NM_133758 USP48 BC021769 USP49 NM_198421 USP50 NM_029163 USP51 XM_982160 USP52 NM_133992 USP53 NM_133857 USP54 AK047620 CYLD NM_173369

* CYLD: cylindromatosis

Example  4. Reverse transcription  Polymerase chain reaction

Insulin isolated from Example 2 was treated to reverse transcriptase polymerase chain reaction using mRNA and primer set prepared in Example 3. The polymerase chain reaction was performed for 5 minutes at 97 ° C. for hot start, and then 30 cycles were performed for 40 seconds at 94 ° C., 40 seconds at 46 ° C., and 40 seconds at 72 ° C., and then extended for 5 minutes at 72 ° C. Rough The amplified polymerase chain reaction product was isolated on 0.8% agarose gel and stained with ethidium bromide.

The result of the reverse transcription polymerase chain reaction is shown in FIG. 1. As can be seen in FIG. 1, USP7 (NM_001003918), USP10 (NM_009462), USP11 (NM_145628), USP16 (NM_024258), USP18 (NM_011909), USP20 (NM_028846), USP22 (NM_001004143), USP25 (NM_013918) (BC088733), USP36 (XM_126772), USP38 (NM_027554), USP39 (NM_138592), USP46 (NM_177561), USP47 (NM_133758), USP53 (NM_133857), and USP54 (AK047620) expression of significantly reduced deubiquitination enzymes It was.

Example  5. Real Time Polymerase Chain Reaction

Real-time polymerase chain reaction was performed to quantitatively quantify the results of reverse transcription polymerase chain reaction analysis obtained in Example 4. Real time polymerase chain reaction was carried out using primers of deubiquitination enzyme showing a total of 16 expression differences obtained in Example 4. The real-time polymerase chain reaction was performed for 40 cycles of 40 seconds at 94 ° C, 40 seconds at 51 ° C, and 40 seconds at 72 ° C. After real-time polymerase chain reaction, the specificity of the product was examined by melting curve analysis. Β-actin (forward primer 5′-CTCTAGACTTCGAGCAGGAG-3 ′, reverse primer 5′-TAGGAGCCAGAGCAGTAATC-3 ′) was used for standardization of the amount.

Real time polymerase chain reaction results are shown in FIG. As can be seen in Figure 2, the de-ubiquitination enzyme, that is, USP53, the relative expression level in the fat precursor cells by insulin treatment was reduced by about 10% than that in the non-insulin-treated fat precursor cells, USP22, 25, 28, 38, 39, 47, and 54 decreased the expression level by about 30% by insulin treatment. In addition, USP10, 16, 18, 46 reduced the expression level by about 40% by insulin treatment. In particular, USP7, 11, 20, 36 can be seen that the expression level in insulin-treated adipocytes more than doubled.

Obesity or diabetes can be diagnosed by measuring the difference in expression of the deubiquitase enzyme genes of SEQ ID NOS: 1-16. Thus, probes and primers obtained from the deubiquitase enzyme genes of SEQ ID NOS: 1-16 can be usefully used to diagnose obesity or diabetes.

Attach sequence list electronic file

Claims (13)

delete delete delete delete delete SEQ ID NOs: 17 and 18; SEQ ID NOs: 19 and 20; SEQ ID NOs: 21 and 22; SEQ ID NOs: 23 and 24; SEQ ID NOs: 25 and 26; SEQ ID NOs: 27 and 28; SEQ ID NOs: 29 and 30; SEQ ID NOs: 31 and 32; SEQ ID NOs: 33 and 34; SEQ ID NOs: 35 and 36; SEQ ID NOs: 37 and 38; SEQ ID NOs: 39 and 40; SEQ ID NOs: 41 and 42; SEQ ID NOs: 43 and 44; SEQ ID NOs: 45 and 46; And a pair of amplification primers for diagnosing obesity or diabetes, characterized in that it is selected from the group consisting of SEQ ID NOs: 47 and 48. The method of claim 6, wherein the primer pair is SEQ ID NOs: 17 and 18; SEQ ID NOs: 21 and 22; SEQ ID NOs: 27 and 28; And amplification primer pairs for diagnosing obesity or diabetes, which are selected from the group consisting of SEQ ID NOs: 35 and 36. delete delete delete A composition for diagnosing obesity or diabetes comprising a primer pair prepared from a gene encoding a deubiquitination enzyme selected from the group consisting of polynucleotides of SEQ ID NOs: 1-16. The method of claim 11, wherein the primer pairs are SEQ ID NOs: 17 and 18; SEQ ID NOs: 19 and 20; SEQ ID NOs: 21 and 22; SEQ ID NOs: 23 and 24; SEQ ID NOs: 25 and 26; SEQ ID NOs: 27 and 28; SEQ ID NOs: 29 and 30; SEQ ID NOs: 31 and 32; SEQ ID NOs: 33 and 34; SEQ ID NOs: 35 and 36; SEQ ID NOs: 37 and 38; SEQ ID NOs: 39 and 40; SEQ ID NOs: 41 and 42; SEQ ID NOs: 43 and 44; SEQ ID NOs: 45 and 46; And SEQ ID NOs: 47 and 48, wherein the composition for diagnosing obesity or diabetes. The method of claim 12, wherein the primer pair is SEQ ID NOs: 17 and 18; SEQ ID NOs: 21 and 22; SEQ ID NOs: 27 and 28; And SEQ ID NOs: 35 and 36, a composition for diagnosing obesity or diabetes.

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