BMB Reports

Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Construction of Yeast Vectors Potentially Useful for Expression of Eukaryotic Genes as ${\beta}$-galactosidase Fusion Proteins

  • Chung, Kyung-Sook (Cell Cycle and Signal Research Unit, Korea Research Institute of Bioscience and Biotechnology, KIST) ;
  • Choi, Won-Ja (Dept. of Biology, Ewha Womans University) ;
  • Lee, Hee-Won (Cell Cycle and Signal Research Unit, Korea Research Institute of Bioscience and Biotechnology, KIST) ;
  • Kim, Kyu-Won (Dept. of Molecular Biology, Pusan National University) ;
  • Yoo, Hyang-Sook (Cell Cycle and Signal Research Unit, Korea Research Institute of Bioscience and Biotechnology, KIST)
  • Received : 1996.03.19
  • Published : 1996.07.31
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Abstract

By both in vitro hydroxylamine mutagenesis of the wild type 3-phosphoglycerate kinase gene (PGK) promoter DNA and insertion of the leu2-d gene, we have created yeast expression vectors potentially useful for production of eukaryotic genes in yeast. The guanine (G) to adenine (A) change at the -3 position from the ATG start codon of the PGK promoter-based vector rendered a 6~7 times elevated expression of the adjacent eukaryotic gene, and insertion of the leu2-d gene in the vector containing the mutated PGK promoter further enhanced the expression of the gene. When expression of the AIDS virus HIV1-gagP17 gene in a lacZ fusion form was examined with this new vector, a 15 times higher level of expression than that from the original PGK promoter was observed. Northern and Southern analysis showed that this elevated expression is due to the production of a high copy number of mRNA by leu2-d gene functioning and by efficient translation of the produced mRNA. Thus, the vector that contained the A at the -3 position from the ATG start codon in the promoter region and the leu2-d gene shows increased expression capability and will be potentially useful for production of eukaryotic genes in yeast.

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