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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
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- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제14권 제3호
- 발행연도
- 2004.1
- 수록면
- 576 - 583 (8page)
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GERI-155 is a macrolide antibiotic containingtwo deoxyhexose molecules and shows antimicrobial activitiesagainst Gram-positive bacteria. Deoxysugar biosyntheticgene cluster of GERI-155 from Streptomyces sp. GERI-155genome was cloned. Four orfs were identified and a putativeorf presumed to be the dTDP glucose-4,6-dehydratase gene wasdesignated as gerE. GerE was expressed in E. coli by using arecombinant expression vector pHJ1. The expressed protein waspurified from E. coli cell lysate by using ammonium sulfatefractionation, and DEAE-sepharose CL-6B and hydroxylapatitecolumn chromatography. The molecular mass of the expressedprotein correlated with the predicted mass that was deducedfrom the cloned gene sequence data. The recombinant proteinwas a homodimer with a subunit relative molecular weight of39,000 Dalton. It was found to have dTDP-glucose 4,6-dehydratase activity and also found to be highly specific fordTDP-glucose as a substrate. The values of Km and Vmax fordTDP-glucose were 32 mM and 335 nmol min-1 (mg protein)-1,respectively. dTTP and dTDP were strong inhibitors of theprotein. NAD+, the coenzyme for dTDP-glucose 4,6-dehydratase,was tightly bound to the expressed protein.
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참고문헌 (5)
참고문헌 신청
Bechthold, A / 1995 / Identification of Streptomyces violeceoruberT22 genes involved in the biosynthesis of granaticin 247 : 610 ~ 620
Bibb / 1984 / Therelationship between base composition and codon usage inbacterial genes and its use for the simple and reliableidentification of protein-coding sequences : 1
Bierman / 1992 / Plasmid cloning vectors for theconjugal transfer of DNA from Escherichia coli toStreptomyces spp se : 43 ~ 49
Bradford / 1976 / A rapid and sensitive method forquantitation of microgram quantities of protein utilizing theprinciple of protein-dye binding : 248 ~ 254
Fernandez, E / 1998 / Identificationof two genes from Streptomyces argillaceus encodingglycosyltransferases involved in transfer of a disaccharideduring biosynthesis of the a
Bibb / 1984 / Therelationship between base composition and codon usage inbacterial genes and its use for the simple and reliableidentification of protein-coding sequences : 1
Bierman / 1992 / Plasmid cloning vectors for theconjugal transfer of DNA from Escherichia coli toStreptomyces spp se : 43 ~ 49
Bradford / 1976 / A rapid and sensitive method forquantitation of microgram quantities of protein utilizing theprinciple of protein-dye binding : 248 ~ 254
Fernandez, E / 1998 / Identificationof two genes from Streptomyces argillaceus encodingglycosyltransferases involved in transfer of a disaccharideduring biosynthesis of the a
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