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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제24권 제3호
- 발행연도
- 2014.1
- 수록면
- 337 - 345 (9page)
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초록· 키워드
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Pichia pastoris is one of the most widely used expression systems for the secretory expressionof recombinant proteins. The secretory expression in P. pastoris usually makes use of the preproMATα sequence from Saccharomyces cerevisiae, which has a dibasic amino acid cleavagesite at the end of the signal sequence. This is efficiently processed by Kex2 protease, resultingin the secretion of high levels of proteins to the medium. However, the proteins that arehaving the internal accessible dibasic amino acids such as KR and RR in the coding regioncannot be expressed using this signal sequence, as the protein will be fragmented. We haveidentified a new signal sequence of 18 amino acids from a P. pastoris protein that can secreteproteins to the medium efficiently. The PMT1-gene-inactivated P. pastoris strain secretes a ~30kDa protein into the extracellular medium. We have identified this protein by determining itsN-terminal amino acid sequence. The protein secreted has four DDDK concatameric internalrepeats. This protein was not secreted in the wild-type P. pastoris under normal cultureconditions. We show that the 18-amino-acid signal peptide at the N-terminal of this protein isuseful for secretion of heterologous proteins in Pichia.
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참고문헌 (22)
참고문헌 신청
Cereghino GPL / 2002 / Production of recombinant proteins in fermenter cultures of the yeast Pichia pastoris / Curr. Opin. Biotechnol 13 : 329 ~ 332
Cereghino JL / 2000 / Heterologous protein expression in the methylotrophic yeast Pichia pastoris / FEMS Microbiol. Rev 24 : 45 ~ 66
Emanuelsson O / 2007 / Locating proteins in the cell using TargetP, SignalP and related tools / Nat. Protoc 2 : 953 ~ 971
Govindappa N / 2011 / Novel prolipase-bovine trypsinogen fusion proteins / Patent No. WO/2011/030347 2011
Govindappa N / 2013 / PMT1gene plays a major role in O-mannosylation of insulin precursor in Pichia pastoris / Protein Expr. Purif 88 : 164 ~ 171
Cereghino JL / 2000 / Heterologous protein expression in the methylotrophic yeast Pichia pastoris / FEMS Microbiol. Rev 24 : 45 ~ 66
Emanuelsson O / 2007 / Locating proteins in the cell using TargetP, SignalP and related tools / Nat. Protoc 2 : 953 ~ 971
Govindappa N / 2011 / Novel prolipase-bovine trypsinogen fusion proteins / Patent No. WO/2011/030347 2011
Govindappa N / 2013 / PMT1gene plays a major role in O-mannosylation of insulin precursor in Pichia pastoris / Protein Expr. Purif 88 : 164 ~ 171
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