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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
- 563 - 569 (7page)
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초록· 키워드
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The DNA sequence of the chitosanase gene(choK) from b-Proteobacterium KNU3 showed an 1,158-bpopen reading frame that encodes a protein of 386 amino acidswith a novel 74 signal peptide. The degenerated primersbased on the partial deduced amino acid sequences fromMALDI-TOF MS analyses yielded the 820 bp of the PCRproduct. Based on this information, double inverse PCRcloning experiments, which use the two specific sets of PCRprimers rather than single set primers, identified the unknown1.2 kb of the choK gene. Subsequently, a 1.8 kb of full choKgene was cloned from another PCR cloning experimentand it was then subcloned into pGEM T-easy and pUC18vectors. The recombinant E. coli clone harboring recombinantpUC18 vector produced a clear halo around the colony in theglycol chitosan plates. The recombinant ChoK protein wassecreted into medium in a mature form while the intracellularChoK was produced without signal peptide cleavage. Theactivity staining of PAGE showed that the recombinantChoK protein was identical to the chitosanase of wildtype.The comparison of deduced amino acid sequences ofchoK revealed that there is 92% identity with that ofSphingobacterium multivorum chitosanase. Judging from theconserved module in other bacterial chitosanases, chitosanaseof KNU3 strain (ChoK) belongs to the family 80 of glycosidehydrolases.
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참고문헌 (7)
참고문헌 신청
Fenton / 1981 / Purification andmode of action of a chitosanase from Penicilliumislandicum : 151 ~ 165
Hoe, H. S / 2002 / Cloning expressionof the gene for inorganic pyrophosphatase of Thermuscaldophilus GK24 and properties of the enzyme 12 : 301 ~ 305
Hong / 2003 / Cloning and characterization of a bifunctional cellulosechitosanasegene from B. licheniformis NBL420 13 : 35 ~ 42
Isabelle / 1992188-193 / Purification andcharacterization of a chitosanase from Streptomyces N174
Jung, W. J / 2002 / Effect of chitinase
Hoe, H. S / 2002 / Cloning expressionof the gene for inorganic pyrophosphatase of Thermuscaldophilus GK24 and properties of the enzyme 12 : 301 ~ 305
Hong / 2003 / Cloning and characterization of a bifunctional cellulosechitosanasegene from B. licheniformis NBL420 13 : 35 ~ 42
Isabelle / 1992188-193 / Purification andcharacterization of a chitosanase from Streptomyces N174
Jung, W. J / 2002 / Effect of chitinase
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