• Title/Summary/Keyword: Gene distribution

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Clonal Analysis of Methicillin-Resistant Staphylococcus aureus Strains in Korea

  • Kim, Jung-Min;Seol, Sung-Yong;Cho, Dong-Taek
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.3
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    • pp.215-224
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    • 2000
  • In this study, the distribution of the mec regulator genes and the presence of the mutation in mecI gene and mec promoter region among 50 MRSA clinical isolates derived from a single university hospital in Korea were analyzed. Among 50 MRSA strains, 13 strains had a deletion of mecI gene, and 37 strains were found to have mutations in mecI gene or mecA promoter region corresponding to a presumptive operator of mecA, i.e., the binding site of the repressor protein. Furthermore, in order to track the evolution of methicillin-resistant Staphylococcus aureus (MRSA) distributed in Korea, we determined the MRSA clonotype by combined use of genetic organization patterns of mec regulator genes, ribotype, and coagulase type. As the result, 48 of 50 MRSA strains could be classified into four distinct clones. Clonotype I is characterized by the coagulase type 3, deletion of mecI gene, and ribotype 1 shared by NCTC10442, the first reported MRSA isolate in England (9 strains). Clonotype II is characterized by the coagulase type 4, C to T substitution at position 202 of mecI gene, and ribotypes 2, 3 and 4 shared by 85/3619 strain isolated in Austria (10 strains). Clonotype III is characterized by the coagulase type 2, mutations of mecA promoter region and/or mecI, and ribotypes 4, 5, and 6 shared by N315 strain isolated in Japan (25 strains). Clonotype IV is characterized by the coagulase type 4, deletion of mecI gene, and ribotype 7 (4 strains). The clonality of two strains could not be determined due to their undefined ribotype.

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Characterization of porcine cytokine inducible SH2-containing protein gene and its association with piglet diarrhea traits

  • Niu, Buyue;Guo, Dongchun;Liu, Zhiran;Han, Xiaofei;Wang, Xibiao
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.12
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    • pp.1689-1695
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    • 2017
  • Objective: The cytokine inducible SH2-containing protein (CISH), which might play a role in porcine intestine immune responses, was one of the promising candidate genes for piglet anti-disease traits. An experiment was conducted to characterize the porcine CISH (pCISH) gene and to evaluate its genetic effects on pig anti-disease breeding. Methods: Both reverse transcription polymerase chain reaction (RT-PCR) and PCR were performed to obtain the sequence of pCISH gene. A pEGFP-C1-CISH vector was constructed and transfected into PK-15 cells to analysis the distribution of pCISH. The sequences of individuals were compared with each other to find the polymorphisms in pCISH gene. The association analysis was performed in Min pigs and Landrace pigs to evaluate the genetic effects on piglet diarrhea traits. Results: In the present research, the coding sequence and genomic sequence of pCISH gene was obtained. Porcine CISH was mainly localized in cytoplasm. TaqI and HaeIII PCR restriction fragment length polymorphism (RFLP) assays were established to detect single nucleotide polymorphisms (SNPs); A-1575G in promoter region and A2497C in Intron1, respectively. Association studies indicated that SNP A-1575G was significantly associated with diarrhea index of Min piglets (p<0.05) and SNP A2497C was significantly associated with the diarrhea trait of both Min pig and Landrace piglets (p<0.05). Conclusion: This study suggested that the pCISH gene might be a novel candidate gene for pig anti-disease traits, and further studies are needed to confirm the results of this preliminary research.

Gene cloning, tissue distribution, and its characterization of Ca2+-activated Cl- channel activated by ginsenosides in Xenopus laevis oocytes (Xenopus laevis oocytes에서 진세노사이드에 의하여 활성화되는 Ca2+-activated Cl- 이온 통로의 유전자 클로닝, 조직 분포 및 채널 특성)

  • Jeong, Sang-Min;Lee, Jun-Ho;Yoon, In-Soo;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.29 no.4
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    • pp.167-175
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    • 2005
  • The $Ca^{2+}-activated$ chloride channel (CLCA) was activated by ginseng total saponin (GTS) in Xenopus oocytes. The reverse transcription PCR (RT-PCR) method was performed with gene specific primers on oocytes. The gene specific primers were deduced from spleen cDNA in expressed sequence tags (EST) database showing high homology to the mouse CLCA. Full length of cDNA sequence was completed by linkage of several 5' and 3'-half cDNA fragments have been sequenced. We named the full cDNA to oCLCA transiently. The oCLCA gene encodes a protein of 911 amino acids with $48.9\%$ identity overall to that of mouse CLCA (mCLCA4). A predicted oCLCA amino acids sequence shows the molecular weight of 108 kDa and has four or more transmembrane domains, and also the one hydrophobic C­terminal domain. oCLCA gene was expressed ubiquitously in various tissues included oocytes, also interfered in oocytes by siRNA for oCLCA. Here, we suggest that oCLCA is a endogenous chloride channel gene in oocytes. We are studying for the identification of oCLCA gene and further physiological research.

The polymorphism of Theileria buffeli major surface protein associate with their clinical signs in holstein in Korea (국내 젖소에서 Theileria buffeli 주요 표면 단백질 유전자의 다양성 분석)

  • Yu, Do-Hyeon;Li, Ying-Hua;Chae, Joon-Seok;Park, Jin-Ho
    • Korean Journal of Veterinary Research
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    • v.51 no.2
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    • pp.107-115
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    • 2011
  • Theileria (T.) buffeli (formerly T. sergenti/T. orientalis) is the major hemo-protozoan distributed in the Far East Asian countries such as Korea, China and Japan. It is responsible for the clinical symptoms of anorexia, ateliosis, anemia, fever and icterus. It also causes abortion and sudden death under severe cases, resulting in economic losses for many livestock farms. The objective of this study was to analyze the genetic diversity of the major surface protein (Msp) gene in T. buffeli in Holstein in Korea, and we characterized the association of the diversification of the Msp gene and its relationship with the pathogenicity of Theileria. For this, complete blood counts and Theileria PCR sequence analysis were performed from 57 Holstein in Jeju Island. A total of 26 PCR positive Holstein (16 anemic and 10 non-anemic) were then randomly selected based on 18s rRNA sequence typing of the Theileria Msp gene. The DNA sequence of the T. buffeli Msp gene in Holstein showed 99.0%, 99.2%, 99.9%, 99.5%, 98.7%, 98.4% and 98.4% homology with T. sergenti, Theileria spp., T. sergenti, Theileria spp., Theileria spp., Theileria spp. and Theileria spp., respectively. The result showed a genetic variation of 57.7% (type I), 3.8% (type II), 15.4% (type III), 7.7% (type IV), 13.5% (type V) and 1.9% (type VI). Type I is the most frequent type in both anemic and non-anemic Holstein while type II was found in only non-anemic Holstein. This results of our study help confirm the diversity of Msp gene types and demonstrate that the gene type distribution of Msp genes varies among Theileria-infected Holstein in Jeju Island.

Genetic Variants of Serum Proteins and Enzymes in Holstein-Friesian Cattle (홀스타인종 유우의 혈청단백질 및 효소의 유전적 변이체)

  • Sang, Byung Chan;Ryoo, Seung Heui;Seo, Kil Woong;Lee, Chang Soo
    • Korean Journal of Agricultural Science
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    • v.22 no.2
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    • pp.163-169
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    • 1995
  • This study was carried out to examine the genetic constitution of serum proteins and enzymes in Holstein Friesian cattle population. The genetic variants of post-transferrin-2(pTf-2), transferrin(Tf), post-albumin(pAlb), ceruloplasmin(Cp) and amylase-I(Am-I) were analyzed by using PAGE(polyacrylamide gel electrophoresis) and STAGE(starch gel electrophoresis). In serum proteins, the pTf-2 locus were observed to be controlled by codominant alleles designated F and S, and the distribution of genotypes were 76.34, 14.50 and 9.10% for pTf-2 FF, FS and SS types, respectively. The gene frequencies of the pTf-2 F and S allele were 0.836 and 0.164. The Tf locus were found to be controlled by four alleles, Tf A, D1, D2 and E at a single locus, and the distribution of genotypes were 6.11, 32.06, 19.08, 1.53, 10.69, 18.32, 9.92 and 2.29% for Tf AA, AD1, AD2, AE, D1D1, D1D2, D2D2 and D2E type, respectively. The gene frequencies of the Tf A, D1, D2 and E wee 0.321, 0.359, 0.298 and 0.019. The pAlb locus were identified to be genetically controlled by two alleles, pAlb F and S allele, and the distribution of genotypes were 32.06, 29.77 and 38.17% for pAlb FF, FS and SS types, respectively. The gene frequencies of the pAlb F and S allele were 0.461 and 0.531. The Alb locus were observed to be controlled by Alb A and B allele, and the gene frequencies of these were 0.996 and 0.004. In serum enzymes, the Cp locus were found to be controlled by F and S allele, and the distribution of genotypes were 46.57, 27.48 and 25.95% for Cp FF, FS and SS types, respectively. The gene frequencies of F and S allele were 0.603 and 0.394. The Am-I locus were observed to be controlled by Am-I B and C allele, and the distribution of genotypes were 39.69, 21.73 and 38.93% for Am-I BB, BC and CC types, the gene frequencies of Am-I B and C were 0.503 and 0.497, respectively.

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A Comparative Study of Parametric Methods for Significant Gene Set Identification Depending on Various Expression Metrics (유전자 발현 메트릭에 기반한 모수적 방식의 유의 유전자 집합 검출 비교 연구)

  • Kim, Jae-Young;Shin, Mi-Young
    • Journal of KIISE:Software and Applications
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    • v.37 no.1
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    • pp.1-8
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    • 2010
  • Recently lots of attention has been paid to gene set analysis for identifying differentially expressed gene-sets between two sample groups. Unlike earlier approaches, the gene set analysis enables us to find significant gene-sets along with their functional characteristics. For this reason, various novel approaches have been suggested lately for gene set analysis. As one of such, PAGE is a parametric approach that employs average difference (AD) as an expression metric to quantify expression differences between two sample groups and assumes that the distribution of gene scores is normal. This approach is preferred to non-parametric approach because of more effective performance. However, the metric AD does not reflect either gene expression intensities or variances over samples in calculating gene scores. Thus, in this paper, we investigate the usefulness of several other expression metrics for parametric gene-set analysis, which consider actual expression intensities of genes or their expression variances over samples. For this purpose, we examined three expression metrics, WAD (weighted average difference), FC (Fisher's criterion), and Abs_SNR (Absolute value of signal-to-noise ratio) for parametric gene set analysis and evaluated their experimental results.

Analysis of Molecular Epidemiological Properties of Staphylococcus aureus Isolates from Domestic Animals and Human Patients by PCR (Polymerase Chain Reaction을 활용한 국내 동물과 사람환자에서 분리한 Staphylococcus aureus 분리주의 분자역학적 특성분석)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.24-37
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    • 2005
  • This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains $(100\%)$ tested and followed by coa-gene $(87.9\%)$, spa-gene $(91.4\%)$ and aroA-gene $(26.7\%)$, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains $(12.1\%)$ were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.

IL-10 gene promoter polymorphisms in Korean gener-alized aggressive periodontitis patients (한국인 전반적 급진성 치주염 환자에서 IL-10 promoter 유전자 다변성에 관한 연구)

  • Ryn, Ji-Sun;Kim, Ok-Su
    • Journal of Periodontal and Implant Science
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    • v.37 no.3
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    • pp.563-573
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    • 2007
  • Genetic polymorphisms associated with aggressive periodontitis have previously been reported. Interleukin-10 is an immunoregulatory cytokine that plays a role in the pathogenesis of periodontitis. Individual capacity for IL-10 production appears to be under genetic influence, The aim of present investigation was to explore possible genetic association of IL-10 gene promoter polymorphisms with generalized aggressive periodontitis. The study population consisted of 37 generalized aggressive periodontitis patients from the Department of Periodontology, Chonnam National University Hospital and 27 control subjects, all the subjects were non-smokers, Genomic DNA was obtained from buccal swab. The IL-10promoter -597, -824, -1082 positions were genotyped by amplifying the polymorphic regions using polymerase chain reaction (PCR) , followed by restriction enzyme digestion and gel electrophoresis. IL-10-597 C (allele 1) to A (allele 2) and IL-10-824 C (allele 1) to T (allele 2) and IL-10-1082 G (allele 1) to A (allele 2) polymorphisms were examined. The results were as follows. 1. In patients, the distribution of genotypes C/C, C/A and NA at Il-10-597 was determined to be 13.5%, 37.8% and 48.7%, respectively and the distribution of genotypes at IL-10-824 was the same as that of IL-10-597. The distribution of genotypes G/G, G/A and NA at IL-10-1082 was found to be 2.7%, 16.2% and 81. 4%, respectively. No statistical difference in genotype distribution was found between the patient and control groups. 2. Allele 2 carriage rate at the three position of the IL-10 promoter region was higher in the control group than the patient group. 3. Allele 2 frequencies at IL-10-597 and -824 positions were higher in female group than male group and its difference was statistically significant(p<0.05). No significant difference in genotype distribution between the control and patient groups. Allele frequency between control and patient groups was not significantly different although allele 2 frequency at the three positions in the IL-10 promoter region appeared to be higher in control group. In conclusion, no clear association between IL-10 gene promoter polymorphisms and generalized aggressive periodontitis in Korean was observed.

The Distribution of C298T Polymorphism in the Oseteocalcin Gene from Korean Male Athletes and its Association with Bone Mineral Density (한국인 남성 운동선수군에서 Osteocalcin 유전자의 C298T 다형성의 분포와 골밀도와의 관계)

  • Jung, In-Geun;Kang, Byung-Yong;Kim, Ji-Young;Oh, Sang-Duk;Ha, Nam-Joo
    • YAKHAK HOEJI
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    • v.50 no.1
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    • pp.26-32
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    • 2006
  • Osteocalcin is a vitamin K dependent and bone specific protein which plays an important role in the regulation of bone and calcium metabolism. In this study, we evaluated the relationship between the C298T polymorphism in the osteocalcin gene and bone mineral density (BMD) in Korean young men and their interaction with physical activity. BMDs of the femoral neck and lumbar spine were measured using dual energy X-ray absorptiometry, and the C298T polymorphism in the osteocalcin gene determined using polymerase chain reaction (PCR)-HindIII restriction fragment length polymorphism (RFLP) method. We did not observe any significant differences in the femoral neck and lumbar spine BMDs across genotypes of this polymorphism in controls, athletes or combined groups, respectively (P>0.05). Therefore, our data suggest that the C298T polymorphism in the osteocalcin gene is not a suitable genetic marker for the susceptibility to BMD.

Cloning and Expression Analysis of Gonadogenesis-associated Gene SPATA4 from Rainbow Trout (Oncorhynchus mykiss)

  • Liu, Bowen;Liu, Shangfeng;He, Shan;Zhao, Ying;Hu, Hongxia;Wang, Zhao
    • BMB Reports
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    • v.38 no.2
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    • pp.206-210
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    • 2005
  • Gonadogenesis is a complicated process which involves multi-gene interactions. A rainbow trout (Oncorhynchus mykiss) gene spermatogenesis associated 4 (SPATA4) was cloned and characterized from adult rainbow trout testis. The cDNA sequence of rainbow trout SPATA4 contains an open reading frame of 1, 081 nucleatides encoding a putative protein of 259 amino acids. The putative protein from rainbow trout shares a 76.8% homology with zebrafish SPATA4. No trans-membrane regions or signal peptide were detected using bioinformatics methods. Subcellular localization analysis revealed that rainbow trout SPATA4 was a nuclear protein with highest possibility (39.1%). Multi-tissue reverse transcriptase PCR (RT-PCR) was performed to examine the distribution of rainbow trout SPATA4 in eleven organs of adult rainbow trout. The result demonstrated that this gene express specifically in testis and slight amount of expression was detected in ovary. Further analysis of SPATA4 characterization and function in rainbow trout may provide insight into the understanding of gonadogenesis process.