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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2008.1
- 수록면
- 260 - 267 (8page)
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The choice of expression vector is very important for industrial production of proteins. Therefore, the systematic mining of promoters over a wider range of genetic resource and/or host is required. We previously reported a novel bidirectional reporting system(pBGR) for the isolation of promoters from metagenome and screened useful promoters that functioned constitutively in E. coli under general culture conditions. Among them, three promoter sequences including each upstream region were amplified by PCR and used to construct new expression vectors. To facilitate subcloning, a multi-cloning site was incorporated into the downstream region of the revere primer sequence. At these sites, GFP, esterase and β-glucosidase were subcloned and analyzed the constitutive expression ability of new promoter in terms of protein solubility and expression level. As a result, these vectors expressed the proteins constitutively to a level of 2~3% of the total cell protein in soluble fraction (>80 %). This study suggested that excavation of metagenomic promoters for construction of expression vector in a certain strain could provide a way for the development of the expression systems.
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참고문헌 (24)
참고문헌 신청
Aoki, T. / 2003 / General properties of GFP-display, an electrophoretic analysis for single amino acid changes in target polypeptides / Anal. Biochem : 107 ~ 115
Baneyx, F. / 2004 / Recombinant protein folding and misfolding in Escherichia coli / Nat. Biotechnol 22 : 1399 ~ 1408
Bhandari, P. / 1997 / An Escherichia coli host strain useful for efficient overproduction of cloned gene products with NaCl as the inducer / J. Bacteriol : 4403 ~ 4406
Cardona, S. T. / 2005 / An expression vector containing a rhamnose-inducible promoter provides tightly regulated gene expression in Burkholderia cenocepacia / Plasmid 54 : 21
Choi, G. S. / 2003 / Construction and characterization of a recombinant esterase with high activity and enantio selectivity to (S)- ketoprofen ethyl ester / Protein. Expr. Pu
Baneyx, F. / 2004 / Recombinant protein folding and misfolding in Escherichia coli / Nat. Biotechnol 22 : 1399 ~ 1408
Bhandari, P. / 1997 / An Escherichia coli host strain useful for efficient overproduction of cloned gene products with NaCl as the inducer / J. Bacteriol : 4403 ~ 4406
Cardona, S. T. / 2005 / An expression vector containing a rhamnose-inducible promoter provides tightly regulated gene expression in Burkholderia cenocepacia / Plasmid 54 : 21
Choi, G. S. / 2003 / Construction and characterization of a recombinant esterase with high activity and enantio selectivity to (S)- ketoprofen ethyl ester / Protein. Expr. Pu
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