• Title/Summary/Keyword: LIM gene

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Enhanced Production of Astaxanthin by Metabolically Engineered Non-mevalonate Pathway in Escherichia coli

  • Jeong, Tae Hyug;Cho, Youn Su;Choi, Seong-Seok;Kim, Gun-Do;Lim, Han Kyu
    • Microbiology and Biotechnology Letters
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    • v.46 no.2
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    • pp.114-119
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    • 2018
  • Astaxanthin is one of the major carotenoids used in pigment has a great economical value in pharmaceutical markets, feeding, nutraceutical and food industries. This study was to increase the production of astaxanthin by co-expression with transformed Escherichia coli using six genes involved in the non-mevalonate pathway. Involved in the non-mevalonate biosynthetic pathway of the strain Kocuria gwangalliensis were cloned dxs, ispC, ispD, ispE, ispF, ispG, ispH and idi genes in order to increase astaxanthin production from the transformed E. coli. And co-expression with the genes to compared the amount of astaxanthin production. This engineered E. coli, containing both the non-mevalonate pathway gene and the astaxanthin biosynthesis gene cluster, produced astaxanthin at $1,100{\mu}g/g$ DCW (dry cell weight), resulting in approximately three times the production of astaxanthin.

Expression of Catalase (CAT) and Ascorbate Peroxidase (APX) in MuSI Transgenic Tobacco under Cadmium Stress

  • Kim, Kye-Hoon;Kim, Young-Nam;Lim, Ga-Hee;Lee, Mi-Na;Jung, Yoon-Hwa
    • Korean Journal of Soil Science and Fertilizer
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    • v.44 no.1
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    • pp.53-57
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    • 2011
  • The MuSI is known as a multiple stress resistant gene with several lines. A previous study using RT-PCR showed that the expression of MuSI gene in tobacco plant induced its tolerance to Cd stress. This study was conducted to examine the enhanced Cd tolerance of the MuSI transgenic tobacco plant through germination test and to understand the role of the involved antioxidant enzymes for the exhibited tolerance. Germination rate of MuSI transgenic tobacco was more than 10% higher than that of wild-type tobacco, and seedlings of MuSI transgenic tobacco grew up to 1.6 times larger and greener than seedlings of wild-type tobacco at 200 and 300 ${\mu}M$ Cd. From the third to the fifth day, CAT activities at 100 and 200 ${\mu}M$ Cd and APX activities at 100, 200 and 300 ${\mu}M$ Cd of MuSI transgenic tobacco were up to two times higher than those of wild-type tobacco. MuSI gene is shown to enhance the activities of antioxidant enzymes resulting in higher tolerance to oxidative stress compared with the control plant.

Cloning and Characterization of the Mycobacterium bovis BCG panB Gene Encoding Ketopantoate Hydroxymethyltransferase

  • Kim, Jin-Koo;Kim, Kwang-Dong;Lim, Jong-Seok;Lee, Hee-Gu;Kim, Sang-Jae;Cho, Sang-Hyun;Jeong, Won-Hwa;Choe, In-Seong;Chung, Thi-Wha;Paik, Sang-Gi;Choe, Yong-Kyung
    • BMB Reports
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    • v.34 no.4
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    • pp.342-346
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    • 2001
  • The Mycobacterium bovis BCG panB gene, encoding ketopantoate hydroxymethyltransferase (KPHMT), was cloned from a ${\lambda}gt11$ genomic library and sequenced. The DNA sequence encodes a protein that contains 281 amino acid residues (M, 29,337) with a high similarity to the KPHMTs. Subcloning of a 846 by open reading frame (ORF), but not a 735 by ORF, into the vector pUC19 led to complementation of the panB mutant of Escherichia coli. The BCG pang gene was overexpressed in E. coli and the KPHMT purified to homogeneity The recombinant protein was further confirmed by an enzymatic assay.

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The Possibility of TBC1D21 as a Candidate Gene for Teat Numbers in Pigs

  • Jin, S.;Lee, J.B.;Kang, K.;Yoo, C.K.;Kim, B.M.;Park, H.B.;Lim, H.T.;Cho, I.C.;Maharani, D.;Lee, J.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.26 no.10
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    • pp.1374-1378
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    • 2013
  • Based on a quantitative traits locus (QTL) study using a $F_2$ intercross between Landrace and Korean native pigs, a significant QTL affecting teat numbers in SSC7 was identified. The strong positional candidate gene, TBC1D21, was selected due to its biological function for epithelial mesenchymal cell development. Sequence analysis revealed six single nucleotide polymorphisms (SNPs) in the TBC1D21 gene. Among these, two SNP markers, one silent mutation (SNP01) for g.13,050A>G and one missense mutation (SNP04) for c.829A>T (S277C), were genotyped and they showed significant associations with teat number traits (p value = 6.38E-05 for SNP01 and p value = 1.06E-07 for SNP04 with total teat numbers). Further functional validation of these SNPs could give valuable information for understanding the teat number variation in pigs.

Genetic characterization and phylogenetic analysis of infectious bronchitis virus isolated in Jeonbuk (전북지역 전염성기관지염 바이러스의 유전적 특성)

  • Chu, Keum-Suk;Kim, Ji-Hyun;Lee, Jeong-Won;Choi, Kwang-Lim
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.51-56
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    • 2015
  • Infectious bronchitis virus (IBV) causes an acute and highly contagious viral disease of chicken that is great economic losses to the poultry industry worldwide. Among the IBV structural proteins, the high rate spike glycoprotein S1 gene mutation and antigenic variant strains have been reported in many countries. During the years 2012~2014, 10 IBV strains were isolated from infected chicken farms distributed in provinces of Jeonbuk. Analysis of the S1 gene sequences amplified from 10 isolated strains with QX strains showed nucleotide homologies ranging from 96.5 to 95.4%. Phylogenetic analysis revealed that all strains were clustered into QX-like groups. This study suggests that QX-like IBVs are circulating in commercial chicken farms in Jeonbuk. Therefore, the continuing survellance is significantly important for prevention and control of BIV infection.

Structure Prediction of the Peptide Synthesized with the Nonribosomal Peptide Synthetase Gene from Bradyrhizobium japonicum

  • JUNG BO-RA;LEE YUKYUNG;LIM YOONGHO;AHN JOONG-HOON
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.656-659
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    • 2005
  • Small peptides synthesized by nonribosomal peptide synthetases (NRPSs) genes are found in bacteria and fungi. While some microbial taxa have few, others make a large number and variety. However, biochemical characterization of the products synthesized by NPRS demands a great deal of efforts. Since the completion of genome projects of numerous microorganisms, the numbers of available NRPSs genes are being expanded. Prediction of the peptides encoded by NRPS could save time and efforts. We chose the NRPS gene from Bradyrhizobium japonicum as a model to predict the peptide structure encoded by NRPS genes. Using computational analyses, the domain structure of this gene was defined, and the structure of a peptide synthesized by this NRPS was deduced. It was found that it encoded a tripeptide consisting of proline-serine-phenylalanine. This method would be helpful to predict the structure of small peptides with various NPRS genes from the genome sequence.

Identification of a p-Cresol Degradation Pathway by a GFP-Based Transposon in Pseudomonas and Its Dominant Expression in Colonies

  • Cho, Ah-Ra;Lim, Eun-Jin;Veeranagouda, Yaligara;Lee, Kyoung
    • Journal of Microbiology and Biotechnology
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    • v.21 no.11
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    • pp.1179-1183
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    • 2011
  • In this study, the chromosome-encoded pcuRCAXB genes that are required for p-cresol degradation have been identified by using a newly constructed green fluorescent protein (GFP)-based promoter probe transposon in the long-chain alkylphenol degrader Pseudomonas alkylphenolia. The deduced amino acid sequences of the genes showed the highest identities at the levels of 65-93% compared with those in the databases. The transposon was identified to be inserted in the pcuA gene, with the promoterless gfp gene being under the control of the pcu catabolic gene promoter. The expression of GFP was positively induced by p-cresol and was about 10 times higher by cells grown on agar than those in liquid culture. In addition, p-hydroxybenzoic acid was detected during p-cresol degradation. These results indicate that P. alkylphenolia additionally possesses a protocatechuate ortho-cleavage route for p-cresol degradation that is dominantly expressed in colonies.

Toxicogenomics Study on ${\alpha}-Naphthylisothiocyanate\;(ANIT)$ Induced Hepatotoxictiy in Mice

  • Hwang, Ji-Yoon;Lim, Jung-Sun;Jeong, Sun-Young;Park, Han-Jin;Cho, Jae-Woo;Yoon, Seok-Joo
    • Molecular & Cellular Toxicology
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    • v.2 no.1
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    • pp.48-53
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    • 2006
  • [ ${\alpha}-Naphthylisothiocyanate$ ] (ANIT) induces intrahepatic cholestasis, involving damage to biliary epitheial cells. This study investigates hepatic gene expression and histopathological alterations in response to ANIT treatment in order to elucidate early time response of ANIT-induced hepatotoxicity. ANIT was treated with single dose (3, 6, and 60 mg/kg) in corn oil by oral gavage. Serum biochemical and histopathological observation were performed for evaluation of hepatotoxicity level. Affymetrix oligo DNA chips were used for gene expression profile by ANIT-induced hetpatoxicity. Hepatic enzyme levels (ALT, AST, and ALP) were increased in 24 hr high dose group. In microscopic observations, moderate hepatocellular necrosis, were confirmed 24 hr high dose groups. We found that gene expression patterns were dependent on time and dose. Our selected genes were related inflammation and immunomodulation. In this study, ANIT-induced hepatotoxicity was involved in acute phase responses and provides evidence for role of neutrophil could be mechanism associated with ANIT-mediated hepatotoxicity.

A Case of Galactosemia with Novel Mutation in the GALT Gene (새로운 GALT 유전자의 돌연변이에 의한 갈락토스혈증)

  • Kim, Shin Ah;Shin, Young Lim;Hong, Yong Hee
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.13 no.2
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    • pp.126-130
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    • 2013
  • Galactosemia is a metabolic disorder inherited by the recessive autosome, and appears by the deficiency of one enzyme out of GALT (Galactose-1-Phosphate Uridyltransferase), GALK (galactokinase), and GALE (epimerase) enzymes, among which the GALT deficiency disease is denominated as classical galactosemia and known to have symptoms such as severe nausea, jaundice, hepatomegaly, sucking difficulty and so on. We report the case of a 16-day-old female baby with the new p.A101D mutation together with p.N413d in the GALT gene analysis found in the neonatal screening test and diagnosed to have galactosemia by the GALT deficiency through the enzyme analysis. For the prognosis prediction, the treatment, the genetic counseling and the prenatal diagnosis of the patients, more detailed genetic diagnosis is required by performing GALT gene analysis, and it is deemed to be necessary to analyze the correlation between the phenotype and the genotype of the domestic galactosemia patients.

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Rapid Enumeration of Listeria monocytogenes in Pork Meat Using Competitive PCR

  • Lim, Hyung-Kun;Hong, Chong-Hae;Choi, Weon-Sang
    • Food Science and Biotechnology
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    • v.14 no.3
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    • pp.387-391
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    • 2005
  • Competitive polymerase chain reaction (cPCR) was used to develop a direct enumeration method of Listeria monocytogenes in pork meat. Pork meat was artificially inoculated with L. monocytogenes and DNA was extracted using guanidine thiocyanate-phenol-chloroform and subjected to PCR amplification. Sixteen primer sets for L. monocytogenes hlyA gene were tested for sensitive detection and the DG69/DG74 primer set was selected. The detection limit achieved with this primer set was as low as 860 colony-forming units (cfu) per 0.1 g of pork meat. When the samples were cultured at $30^{\circ}C$ for 16 hr in Brain Heart Infusion (BHI) medium, even a single bacterium could be detected with this primer set by PCR. For cPCR, the hlyA gene, which features a 148 bp-deletion, was cloned in the pGEM-4Z vector. A known amount of competitor DNA which has the same primer binding sites was co-amplified with L. monocytogenes total DNA from the artificially inoculated pork meat. The cell-number determined by cPCR was approximately equal to cfu from the Most Probable Number (MPN) method. The whole procedure took only 5 hr.