• Title/Summary/Keyword: DHPLC

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Analysis of Interleukin-6 Gene Polymorphisms in Behcet’s Disease Using RFLP and DHPLC (RFLP와 DHPLC를 이용한 베체트병 환자에서 Interleukin-6 유전자 다형성의 분석)

  • Jang, Won-Cheol;Park, Sang-Beom;Nam, Yun-Hyeong;Lee, Jae-Sik
    • Journal of the Korean Chemical Society
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    • v.50 no.2
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    • pp.123-128
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    • 2006
  • Behcet's disease (BD) is a systemic vasculitis characterized by recurrent oral and genital ulcers, and ocular inflammation, and which may involve the joints, skin, central nervous system and gastrointestinal tract. Although the exact pathogenesis for BD is not completely understood, it has been suggested that the disease is triggered in genetically susceptible individuals by environmental factors, such as microbial agents. It is noted that multiple genes, including MHC (major histocompatibility complex) and non-MHC genes, are implicated in the pathogenesis of BD. This study tries to determine whether IL-6 gene polymorphisms are associated with susceptibility to Behcet's disease in Koreans. Gene polymorphisms were typed by VNTR (variable number of tandem repeat), RFLP (restriction fragment length polymorphism), DHPLC (denaturing high performance liquid chromatography).There were no evidences for genetic association conferred by the IL-6prom polymorphism. However, significant differences in the IL-6vntr genotype and allele frequencies were found between patients with BD and controls. The IL-6vntr*C allele appeared to be an additional susceptibility gene to Korean BD. Further studies in other populations and gene are required to confirm these results.

A Simple Method for the Detection and Identification of Korean Brown Cattle(Hanwoo) using DHPLC

  • Kang, Won;Park, Sane-Bum;Nam, Youn-Hyoung;An, Young-Chang;Lee, Sang-Hyun;Park, Jin-Keun;Lee, Jae-Sik;Jang, On-Cheoul;Park, Su-Min;Park, Yong-Hyun;Ko, Kyung-Hee;Moon, Sung-Sil
    • Proceedings of the Korean Society for Food Science of Animal Resources Conference
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    • 2006.05a
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    • pp.179-183
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    • 2006
  • Denaturing high-performance liquid chromatography(DHPLC) is used in a wide variety of genetic applications and is an efficient method for detection of mutations involving one or a few nucleotides. We developed a high-throughput DHPLC method for identifying polymorphisms in the MC1R gene that are characteristic of Hanwoo cattle. We compared 10 tissue samples from Hanwoo cattle, 10 samples from Holstein cattle and 10 samples from Hanwoox Holstein crossbred cattle to determine whether DHPLC analysis can be used to distinguish between these genotypes. Samples obtained from Hanwoo cattle had a unique profile of peaks that could be used as a molecular fingerprint for this breed. We also analyzed two hundred samples in a trial in which we were blinded to the genotype of the samples and correctly identified the breed-of-origin of 594 out of 600 sequence variations(99%).

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Discrimination of Hanwoo from Holstein and Mixed Beef by DHPLC (변성 고성능 액체 크로마토그래피를 이용한 한우, 젖소 그리고 혼입육의 구분)

  • Ahn, Young-Chang;Cho, Min-Ho;Seo, Jae-Won;Yoon, Il-Kyu;Jung, Duck-Hyun;Lee, Eun-Young;Nam, Youn-Hyoung;Park, Su-Min;Jang, Won-Cheoul
    • Journal of the Korean Chemical Society
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    • v.53 no.6
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    • pp.742-748
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    • 2009
  • In the meat industry, correct breed information in food labeling is required to assure meat quality. Genetic markers provide corroborating evidence to identify breed. We described the development of DNA markers to discriminate between Korean beef cattle (Hanwoo), Holstein, and mixed cow beefs. As most breeds are standardized for coat colour, the melanocortin 1 receptor (MC1R) gene, involved in the regulation of eu/pheomelanins synthesis, has been suggested as marker for breed traceability of products of animal origin. We also designed sex-determining region Y (SRY) gene specific primers for Y chromosome detection. In this study, fragments of MC1R gene and SRY gene were amplified by multiplex-PCR and subjected to digestion by MspA1I restriction endonuclease. Reaction products were analysised by denaturing high performance liquid chromatography (DHPLC). As a result, we identified 6 DHPLC peak types from MC1R gene and SRY gene analysis. DHPLC method showed more sensitive than RFLP method for DNA fragments analysis. Therefore, DHPLC method can apply to identify for Hanwoo, Holstein and mixed beef.

A Study of Tissue-Specific Expression and Analysis of Structure and DNA Sequences of Porcine Cytochrome P450 Aromatase Genes

  • Ghimeray, Amal-Kumar;Chae, Sung-Hwa;Lee, Eun-Ju;Quan Feng;Nina Oriero;Park, Inho
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.196-196
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    • 2004
  • By use of RT-PCR coupled with DHPLC technique (WAVE analysis), pattern of isoforms of porcine cytochrome P450 aromatase gene was investigated. Relatively higher expression of aromatase mRNA was observed in testis than in ovary and this result accounted for the previous findings of higher blood estrogen level in male compared with female in this species. (omitted)

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Development of Clamping Probe for Rare DNA Detection using Universal Primers

  • Kim, Meyong Il;Lee, Ki-Young;Cho, Sang-Man
    • Fisheries and Aquatic Sciences
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    • v.17 no.3
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    • pp.339-344
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    • 2014
  • PCR amplification with universal primer is a useful tool for speciation of symbionts in marine eukaryote coupled with robust separation method such as denaturing high performance chromatography (DHPLC). To overcome the biased amplification, clamping PCR is recommended to suppress the amplification of host gene. In this study, we evaluated the efficiency of rare gene detection for two kinds of clamping probes which were successfully utilized for eukaryotic symbiont analysis: C3 linked nucleotide (C3) and peptide nucleic acid (PNA). PNA was 3-4 orders of magnitude higher than that of C3 tested in clamping efficiency and rare gene detection. This represented that PNA could be a more competent clamping probe for the enhancement of PCR amplification for rare symbiont genes.

Identification of strain harboring both aac(6')-Ib and aac(6')-Ib-cr variant simultaneously in Escherichia coli and Klebsiella pneumoniae

  • Kim, Yun-Tae;Jang, Ji-Hyun;Kim, Hyun-Chul;Kim, Hyo-Gyeong;Lee, Kyoung-Ryul;Park, Kyung-Sun;Lee, Hee-Joo;Kim, Young-Jin
    • BMB Reports
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    • v.44 no.4
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    • pp.262-266
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    • 2011
  • The aac(6')-Ib gene is the most prevalent gene that encodes aminoglycoside-modifying enzymes and confers resistance to tobramycin, kanamycin, and amikacin. The aac(6')-Ib-cr variant gene can induce resistance against aminoglycoside and fluoroquinolone simultaneously. Two main methods, sequence analysis and the restriction enzyme method, can detect the aac(6')-Ib-cr variant in clinical strains. We collected the 85 strains that were believed to be aac(6')-Ib positive from clinical isolates. Among them, 38 strains were the wild-type; the remaining 47 strains were the aac(6')-Ib-cr variant. Of these 47 strains, 19 simultaneously harbored aac(6')-Ib and aac(6')-Ib-cr. Our study aims to report the characteristics of the 19 strains that simultaneously harbored both genes. This study is the first investigation published in Korea of strains that included both aac(6')-Ib and aac(6')-Ib-cr variant.

Expression Characterization, Polymorphism and Chromosomal Location of the Porcine Calsarcin-3 Gene

  • Wang, Heng;Yang, Shulin;Tang, Zhonglin;Mu, Yulian;Cui, Wentao;Li, Kui
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.9
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    • pp.1349-1353
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    • 2007
  • Calcineurin is a calmodulin dependent protein that functions as a regulator of muscle cell growth and function. Agents capable of interacting with calcineurin could have important applications in muscle disease treatment as well as in the improvement of livestock production. Calsarcins comprise a family of muscle-specific calcineurin binding proteins which play an important role in modulating the function of calcineurin in muscle cells. Recently, we described the first two members of the calsarcin family (calsarcin-1 and calsarcin-2) in the pig. Here, we characterized the third member of the calsarcin family, calsarcin-3, which is also expressed specifically in skeletal muscle. However, unlike calsarcin-1 and calsarcin-2, the calsarcin-3 mRNA expression in skeletal muscle kept rising throughout the prenatal and postnatal development periods. In addition, radiation hybrid mapping indicated that porcine calsarcin-3 mapped to the distal end of the q arm of pig chromosome 2 (SSC2). A C/T single nucleotide polymorphism site in exon 5 was genotyped using the denaturing high performance liquid chromatography (DHPLC) method and the allele frequencies at this locus were significantly different among breeds.