• Title/Summary/Keyword: weight encoding

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Cloning and Expression of a Collagenase Gene from the Marine Bacterium Vibrio vulnificus CYK279H

  • Kim, Bong-Jo;Kim, Hak-Ju;Hwang, Sun-Hee;Bae, Seoung-Kwon;Ha, Soon-Duck;Kim, Jong-Deog;Kong, Jai-Yul
    • Journal of Microbiology and Biotechnology
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    • v.8 no.3
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    • pp.245-250
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    • 1998
  • A gene encoding an extracellular collagenase from the marine bacterium Vibrio vulnificus CYK279H was cloned into E. coli JM83 using the multicopy plasmid vector pUC19. The cloned strain of recombinant E. coli showing collagenase activity had an insert fragment of 3.5 kb and was named E. coli JM83/pKCL 279H. The cloned strain produced two different collagenase during cultivation. These enzymes, named collagenase-I and -II, were purified from the culture supernatant. SDS-PAGE indicated that collagenase-I had a molecular weight of 41 kDa and collagenase-II had a weight of 37 kDa. The N-terminal amino acid sequence of collagenase-I from the cloned strain, E. coli JM83/pKCL279H was determined and was not found to be similar to any other known collagenases. The optimum pH and temperature of the purified collagenase-I were 7.8 and $37^{\circ}C$, respectively.

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Effect of Genistein and Soy Protein on Lipids Metabolism in Ovariectomized Rats (난소절제 랫드에서 제니스테인과 콩단백질이 지질 대사에 미치는 영향)

  • Lee Young Min;Jung Myeong Ho;Lee Yeon Sook;Song Jihyun
    • Journal of Nutrition and Health
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    • v.38 no.4
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    • pp.267-278
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    • 2005
  • Postmenopausal women or ovariectomized rats are associated with increased cholesterol levels, which are risk factors of metabolic syndrome and cardiovascular diseases. Increased prevalence of metabolic syndrome after menopause might be associated with estradiol deficiency. Harmful effect of estradiol hampers the casual usage of hormone to prevent the metabolic syndrome. Soy protein has been reported to show several beneficial effects on health, however it is unclear which components of soy protein is responsible for anti-obesity and hypocholesterolemic effects. Soy isoflavones, gem-stein and daizein, are suggested to have anti-obesity and hypocholesterolemic effects but with inconsistency. The present study investigated the effect of supplementation of genistein (experiment I) and soy protein containing isoflavones (experiment II) to high fat diet on body weight gain, food intake, liver and fat tissue weight and the lipid levels in ovariectomized rats. Plasma and hepatic lipid contents and the mRNA levels of genes encoding lipid metabolism related proteins, such as CPT1 and HMGR were measured. Ovariectomy increased body weight, fat tissue weight and plasma and hepatic lipid levels which increase the risk of metabolic syndrome. Soy protein could improve plasma and hepatic lipids levels. Soy protein also increased hepatic CPT1 and HMGR mRNA levels. Plasma and hepatic lipids levels could not be decreased by dietary genistein alone. In contrast, lipids levels could be decreased by isoflavone-fortified soy protein, suggesting that the ingestion of soy protein enriched with isoflavone gives more benefit for protecting postmenopausal women from metabolic syndrome.

Development of transgenic potato with high content of functional carotenoids by using metabolic engineering (대사공학기술을 이용한 기능성 carotenoids 고 생산 감자의 개발 현황)

  • Ahn, Mi-Jeong;Bae, Jung-Myung;Lee, Shin-Woo
    • Journal of Plant Biotechnology
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    • v.37 no.4
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    • pp.388-393
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    • 2010
  • Recently, a number of successful research reports are accumulated to increase the carotenoid level in potato tuber such as $\beta$-carotene, precursor of vitamin A and keto-carotenoid like astaxanthin in which is not synthesized in most plants tissue since it does not contain a specific enzyme to add keto-ring in carotenoid molecule. In particular, keto-carotenoids are more interested due to their strong antioxidant activity. Currently, the content of $\beta$-carotene was increased up to 3,600-fold ($47\;{\mu}g/g$ dry weight) when compared to the control potato tuber, parental cultivar for genetic modification. In addition, astaxanthin, one of the major keto-carotenoid was accumulated up to $14\;{\mu}g/g$ dry weight in potato tuber with red color by over expressing the gene encoding $\beta$-carotene ketolase isolated from marine microorganisms. In this article, we summarized carotenogenesis-related genes that have been used for metabolic engineering of carotenoid biosynthetic pathway in potato. Furthermore, strategies for the accumulation of carotenoids and ketocarotenoids in specific potato tuber, bottle necks, and future works are discussed.

Optimal Design of a Transformer Core Using DEAS (DEAS를 이용한 변압기 코아의 최적설계)

  • Kim, Tae-Gyu;Kim, Jong-Wook
    • The Transactions of The Korean Institute of Electrical Engineers
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    • v.56 no.6
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    • pp.1055-1063
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    • 2007
  • This paper introduces an optimal design technique for a 250-watt isolation transformer using an optimization method, dynamic encoding algorithm for searches (DEAS). Although the optimal design technique for transformers dates back to 1970s and various optimization methods have been developed so far, literature concerning global optimization for transformer core design is rarely found against its importance. In this paper, core configuration of the isolation transformer whose performance is computed by complex mathematical steps is optimized with DEAS. The optimization result confirms that DEAS can be successfully employed to transformer core design for various performance specifications only by adjusting weight factors in cost function.

Pseudoalteromonas carrageenovora 유래 Arylsulfatase의 cloning과 재조합 E. coli에서 과발현

  • Im, Jae-Myeong;Kim, Hyeong-Rak;Kim, Seong-Gu;Nam, Su-Wan
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.571-575
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    • 2003
  • A marine aerobic Gram-negative bacterium, Pseudoalteromonas carrageenovora, has been blown to hydrolyze carrageenans, the sulfated galactans of red algae, and to desulfate oligo kappa-carrageenans. Recently, the gene encoding arylsulfatase (aryl-sulfate sulfohydrolase, E.C.3.1.6.1) of A. carrageenovora was cloned and the nucleotide sequence was reported. Enzymatic hydrolysis of sulfate groups in agaropectin simplifies the process of agarose preparation. In order to overproduce the enzyme, the arylsulfatase gene (astA, 984 bp ORF) from P. carrageenovora genome was amplified by PCR and subcloned into the pET21a vector. When the constructed plasmid pAST-A1 was introduced into E, coli BL21(DE3), the transformant on LB plate containing IPTG showed the hydrolyzing activity for p-nitrophenyl sulfate. Most of arylsulfatase activity was found in the cell lysate, but at $50\;{\sim}\;5000\;{\mu}M$ IPTG concentration the activity was found both in the culture supernatant and the cell lysate. The molecular weight of the recombinant enzyme was estimated to be 34 kDa by SDS-PAGE.

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Cloning and Characterization of a Gene Encoding 22 kDa Functional Protein of Bacteriophage MB78

  • Gupta, Lalita;Chakravorty, Maharani
    • BMB Reports
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    • v.38 no.2
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    • pp.161-166
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    • 2005
  • Functional protein of MB78 bacteriophage having apparent molecular weight of 22 kDa is expressed from 1.7 kb HindIII G fragment. The nucleotide sequence of this fragment showed two open reading frames of 222 and 196 codons in tail-to-tail orientation separated by a 62-nucleotide intercistronic region. The ORF of 22 kDa protein is present in opposite orientation, i.e. in the complementary strand, preceded by a strong ribosomal binding site and a promoter sequence. Another ORF started from the beginning of the fragment whose promoter region and translational start site lies in the 0.45 kb HincII U fragment which is located next to the HindIII G fragment, that has the sequence for DNA bending. 3' end of the fragment has high sequence homology to the EaA and EaI proteins of bacteriophage P22, a close relative of MB78 phage.

Molecular Cloning and mRNA Expression a Glutathione S-Transferase cDNA from the Spider, Araneus ventricosus

  • Shin, Geun Ho;Kim, Hyung Suk;Kwon, Dong Wook;Lee, Jin Young;Byeon, Gyeong Min;Sohn, Hung Dae;Jin, Byung Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.1
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    • pp.65-71
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    • 2004
  • A fat body-specific glutathione S-transferase cDMA was cloned from the spider, Araneus ventricosus. The cDNA encoding A. ventricosus glutathione S-transferase (AvGST) is 645 base pairs long with an open reading frame of 215 amino acid residues with a calculated molecular weight of approximately 24 kDa. Northern blot analysis showed the tissue-specifically expression of AvGST in the A. ventricosus fat body.

Molecular Cloning of the Myosin Light Chain-2 cDNA of Gryllotalpa orientalis

  • Cha, So Young;Hwang, Jae Sam;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.1
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    • pp.127-130
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    • 2004
  • We describe here the cloning and characterization of a cDNA encoding a putative myosin light chain-2 (MLC-2) from the mole cricket, Gryllotalpa orientalis. The G. orientalis MLC-2 cDNA sequences comprised of 615 bp with 205 amino acid residues with a calculated molecular weight of approximately 23 kDa. The deduced protein sequence of G. orientalis MLC-2 cDNA showed 64% and 54% identity to Drosophila melanogaster MLC-2 and D. yakuba MLC-2, respectively. Northern blot analysis confirmed the muscle-specific expression of G. orientalis MLC-2.

Purification and characterization of alcohol dehydrogenase encoded by Zymomonas mobilis gene in Escherichia coli

  • 신병식;윤기홍;박무영
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.521.3-522
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    • 1986
  • A gene encoding alcohol dehydrogenase (ADH) in Zymomonas mobilis was cloned into E. coli JM 83 with plasmid pUC 9. The ADH produced by the E. coli transformant was purified bysonication, (NH$^4$)2SO4 fractionation, Affi-Gel blue and hydroxylapatite chromatography. The ADH produced by Z. mobilis was also purified by the same procedures. The two enzyme preparations were characterized and compared. It was found that the E. coli ADH was identical to one of two ADH isozymes of Z. mobilis. Analytical gel filtrations led to the conclusion that the molecule of E. coli ADH was composedv of four subunits having molecular weight of 40,000 (+1,000) dalton each The effect of metal ions on ADH activity and optimum pH were investigated.

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Alginate Lyase Production of Halophilic Pseudomonas sp. by Recombinant Escherichia coli

  • Kong, In-Soo;Kim, Young-Ok;Kim, Jin-Man;Kim, Sung-Koo;Oh, Doo-Hwan;Yu, Ju-Hyun;Kong, Jal-Yul
    • Journal of Microbiology and Biotechnology
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    • v.5 no.2
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    • pp.92-95
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    • 1995
  • Halophilic Pseudomonas sp.W7 isolated from laver in the southem sea of Korea showed alginate lyase activity. Gene (aly) encoding alginate lyase was cloned in E.coli JM83 and the N-terminal amino acid sequence of the enzyme was determined after purificaion. The recombinant enzyme has been shown to have a molecular weight of about 40kDa after 12% SDS-polyacrylamide gel electrophoresis.

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