• Title/Summary/Keyword: Gene Identification

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Major SNP identification for oleic acid and marbling score which are associated with Korean cattle (한우의 올레인산과 근내지방도에 영향을 미치는 유전자 내 에스엔피 규명)

  • Oh, Dong-Yep;Yeo, Jung-Sou;Lee, Jea-Young
    • Journal of the Korean Data and Information Science Society
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    • v.25 no.5
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    • pp.1011-1024
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    • 2014
  • This study is to identify the relationship between unsaturated fatty acids, which are indicators of beef flavor, and unsaturated fatty acid biosynthetic enzymes, which are associated with SNPs in the SCD, SREBPs, $PPAR{\gamma}$, FABP4, FASN and LPL in Hanwoo population. For analysis of fatty acid in Hanwoo, we used to Hanwoo steer(n=513). Also, following an analysis of the relationship raised from Gyeongbuk province region. FABP4; g.3977-325 T>C was selected and the distribution of beef grade of g.3977-325 T>C related in field trial was proved very excellent.

Identification of Polymorphisms in CYP2E1 Gene and Association Analysis among Chronic HBV Patients

  • Chun, Ji-Yong;Park, Byung-Lae;Cheong, Hyun-Sub;Kim, Jason-Y.;Park, Tae-Joon;Lee, Jin-Sol;Lee, Hyo-Suk;Kim, Yoon-Jun;Shin, Hyoung-Doo
    • Genomics & Informatics
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    • v.7 no.4
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    • pp.187-194
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    • 2009
  • Cytochrome P450 2E1 (CYP2E1) is a member of the cytochrome P450 superfamily, and it is a key enzyme responsible for the metabolic activation of many smallmolecular-weight compounds such as alcohol, which is classified as a human carcinogen. In this study, we identified 19 single nucleotide polymorphisms (SNPs) in CYP2E1 in Korean population. In these SNPs, we examined possible genetic association of CYP2E1 polymorphisms with HBV clearance and the risk of hepatocellular carcinoma (HCC). Five common polymorphic sites were selected, CYP2E1 polymorphisms at rs381-3867, rs3813870, rs2070673, rs2515641 and rs2480257, considering their allele frequencies, haplotype-tagging status and LDs for genotyping in larger-scale subjects (n=1,092). Statistical analysis demonstrated that CYP2E1 polymorphisms and haplotypes show no significant association with HBV clearance, HCC occurrence and onset age of HCC (p>0.05). Previous studies, however, have shown contradictory findings on associations of CYP2E1 polymorphisms with CYP2E1 activities and HCC risk. Comparing the contrasting results of previous researches suggest that CYP2E1 polymorphism is associated with CYP2E1 activity induced by ethanol, but is not directly associated with HCC risk. CYP2E1 variation/haploype information identified in this study will provide valuable information for future studies on CYP2E1.

IVET-based Identification of Virulence Factors in Vibrio vulnificus MO6-24/O

  • Lee, Ko-Eun;Bang, Ji-Sun;Baek, Chang-Ho;Park, Dae-Kyun;Hwang, Won;Choi, Sang-Ho;Kim, Kum-Soo
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.234-243
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    • 2007
  • Vibrio vulnificus is an opportunistic pathogen that causes septicemia in humans. To identify the genes associated with its pathogenicity, in vivo expression technology (IVET) was used to select genes specifically expressed in a host, yet not significantly in vitro. Random lacZ-fusions in the genome of V vulnificus strain MO6-24/O were constructed using an IVET vector, pSG3, which is a suicide vector containing promoterless-aph and -lacZ as reporter genes. A total of ${\sim}18,000$ resulting library clones were then intraperitoneally injected into BALB/c mice using a colony forming unit (CFU) of $1.6{\times}10^6$. Two hours after infection, kanamycin was administered at $200{mu}g$ per gram of mouse weight. After two selection cycles, 11 genes were eventually isolated, which were expressed only in the host. Among these genes, VV20781 and VV21007 exhibiting a homology to a hemagglutinin gene and tolC, respectively, were selected based on having the highest frequency. When compared to wild-type cells, mutants with lesions in these genes showed no difference in the rate of growth rate, yet a significant decrease in cytotoxicity and the capability to form a biofilm.

Identification and Functional Analysis of Vibrio vulnificus SmcR, a Novel Global Regulator

  • Lee, Jeojng-Hyun;Rhee, Jee-Eun;Park, U-Ryung;Ju, Hyun-Mok;Lee, Byung-Cheol;Kim, Tae-Sung;Jeong, Hye-Sook;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.325-334
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    • 2007
  • Recently, quorum sensing has been implicated as an important global regulator controlling the production of numerous virulence factors such as capsular polysaccharides in bacterial pathogens. The nucleotide and deduced amino acid sequences of smcR, a homolog of V. harveyi luxR identified from V. vulnificus ATCC29307, were analyzed. The amino acid sequence of SmcR from V. vulnificus was 72 to 92% similar to those of LuxR homologs from Vibrio spp. Functions of SmcR were assessed by the construction of an isogenic mutant, whose smcR gene was inactivated by allelic exchanges, and by evaluating its phenotype changes in vitro and in mice. The disruption of smcR resulted in a significant alteration in biofilm formation, in type of colony morphology, and in motility. When compared with the wild-type, the smcR mutant exhibited reduced survival under adverse conditions, such as acidic pH and hyperosmotic stress. The smcR mutant exhibited decreased cytotoxic activity toward INT 407 cells in vitro. Furthermore, the intraperitoneal $LD_{50}$ of the smcR mutant was approximately $10^2$ times higher than that of parental wild-type. Therefore, it appears that SmcR is a novel global regulator, controlling numerous genes contributing to the pathogenesis as well as survival of V. vulnificus.

Characterization of Glycerol Dehydrogenase from Thermoanaerobacterium thermosaccharolyticum DSM 571 and GGG Motif Identification

  • Wang, Liangliang;Wang, Jiajun;Shi, Hao;Gu, Huaxiang;Zhang, Yu;Li, Xun;Wang, Fei
    • Journal of Microbiology and Biotechnology
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    • v.26 no.6
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    • pp.1077-1086
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    • 2016
  • Glycerol dehydrogenases (GlyDHs) are essential for glycerol metabolism in vivo, catalyzing its reversible reduction to 1,3-dihydroxypropranone (DHA). The gldA gene encoding a putative GlyDH was cloned from Thermoanaerobacterium thermosaccharolyticum DSM 571 (TtGlyDH) and expressed in Escherichia coli. The presence of Mn2+ enhanced its enzymatic activity by 79.5%. Three highly conserved residues (Asp171, His254, and His271) in TtGlyDH were associated with metal ion binding. Based on an investigation of glycerol oxidation and DHA reduction, TtGlyDH showed maximum activity towards glycerol at 60℃ and pH 8.0 and towards DHA at 60℃ and pH 6.0. DHA reduction was the dominant reaction, with a lower Km(DHA) of 1.08 ± 0.13 mM and Vmax of 0.0053 ± 0.0001 mM/s, compared with glycerol oxidation, with a Km(glycerol) of 30.29 ± 3.42 mM and Vmax of 0.042 ± 0.002 mM/s. TtGlyDH had an apparent activation energy of 312.94 kJ/mol. The recombinant TtGlyDH was thermostable, maintaining 65% of its activity after a 2-h incubation at 60℃. Molecular modeling and site-directed mutagenesis analyses demonstrated that TtGlyDH had an atypical dinucleotide binding motif (GGG motif) and a basic residue Arg43, both related to dinucleotide binding.

Functional Identification and Expression of Indole-3-Pyruvate Decarboxylase from Paenibacillus polymyxa E681

  • Phi, Quyet-Tien;Park, Yu-Mi;Ryu, Choong-Min;Park, Seung-Hwan;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.18 no.7
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    • pp.1235-1244
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    • 2008
  • Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.

Identification of the Precursor for the Soybean Kunitz Trypsin Inhibitor (대두 Kunitz Trypsin Inhibitor 전구체의 동정)

  • Kim, Chung-Ho;Kim, Su-Il;Choi, Yang-Do
    • Applied Biological Chemistry
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    • v.32 no.3
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    • pp.222-231
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    • 1989
  • Three classes of proteinase inhibitors are known in soybean; the Kunitz trypsin inhibitor (SKTI), the Bowman-Birk proteinase inhibitor and its isoinhibitors. To study the molecular structure and expression characteristics of the SKTI, antibody was obtained by immunizing rabbit with the SKTI purified from soybean by preparative electrophoresis. Anti-SKTI antibody was not only specific for mature SKTI in soybean seed but also recognized the precursor which was synthesized in vitro. Translation in vitro was carried out in wheat germ extract with polyadenylated mRNA isolated from developing soybean seeds. One of the seed specific translation products, MW 24K, was identified to be the precursor for the SKTI by immunoprecipitation with anti-SKTI antibody. Mature SKTI of MW 20K, however, was not detected in the translates in vitro. These results suggest that the precursor polypeptide is synthesized from the mRNA and is cleaved to yield mature SKTI in soybean seed. The SKTI gene was expressed with the maturation of soybean seed in a tissue-specific and development stage-specific manner.

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The Effects of Breeding Environment Adjustment in FABP4 Gene Identification of Korean Cattle (한우의 FABP4 유전자 선별에서 사육환경 보정 효과)

  • Kim, Hyun-Ji;Lee, Jea-Young
    • The Korean Journal of Applied Statistics
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    • v.28 no.5
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    • pp.859-870
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    • 2015
  • Economic-traits of livestock are affected by environmental and genetic factors. We are interested in genetic factors that influence the economic-traits of Korean cattle. It is necessary to adjust environmental factors in order to enhance the accuracy of the genetic effect analysis. In this paper, we propose a statistical model of Korean cattle that exclude environmental breeding farm and age factors. We formulated an adjusted economic-trait value, and applied multifactor dimensionality reduction (MDR) method to data of before-and-after adjustment to identify major FABP4 genes. We were able to increase the accuracy of the analysis after adjustment and identify superior FABP4 genes that influence grade and fatty acid.

Culture-Independent Methods of Microbial Community Structure Analysis and Microbial Diversity in Contaminated Groundwater with Major Pollutants (주요 오염물질로 오염된 지하수에서 미생물의 무배양식 군집분석방법과 미생물상에 대한 조사방법 연구)

  • Kim Jai-Soo
    • Journal of Soil and Groundwater Environment
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    • v.11 no.3
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    • pp.66-77
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    • 2006
  • This review inquired the recently applied molecular biological and biochemical methods analyzing the microbial community structure of groundwater and, as a result, summarized the functional or taxonomic groups of active microorganisms with major contaminants in groundwater. The development of gene amplification through PCR has been possible to figure out microbial population and identification. Active microbial community structures have been analyzed using a variety of fingerprinting techniques such as DGGE, SSCP, RISA, and microarray and fatty acid analyses such as PLFA and FAME, and the activity of a specific strain has been examined using FISH. Also, this review included the dominant microflora in groundwater contaminated with fuel components such as n-alkanes, BTEX, MTBE, and ethanol and chlorinated compounds such as TCE, PCE, PCB, CE, carbon tetrachloride, and chlorobenzene.

Identification of 12 radiation-resistant bacterial species in the phylum Proteobacteria new to Korea

  • Han, Joo Hyun;Maeng, Soohyun;Park, Yuna;Lee, Sang Eun;Lee, Byoung-Hee;Lee, Ki-eun;Kim, Myung Kyum
    • Journal of Species Research
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    • v.9 no.2
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    • pp.85-104
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    • 2020
  • In 2019, after a comprehensive investigation of indigenous prokaryotic species in Korea, a total of 12 bacterial strains assigned to the phylum Proteobacteria were isolated from soil. With the high 16S rRNA gene sequence similarity (>98.8%) and formation of a robust phylogenetic clade with the closest species, it was determined that each strain belonged to independent, predefined bacterial species. This study identified two species in the family Burkholderiaceae, one species in the family Comamonadaceae, two species in the family Oxalobacteraceae, one species in the family Micrococcaceae, one species in the family Bradyrhizobiaceae, one species in the family Methylobacteriaceae, one species in the family Rhizobiaceae, one species in the family Rhodocyclaceae, and one species in the family Sphingomonadaceae. There is no official report about these 12 species in Korea, so are described as unreported bacterial species in Korea in this study. Gram reaction, basic biochemical characteristics, colony, and cell morphology are also described in the species description section.