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KMID : 1234420090370040389
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Korean Journal of Microbiololgy and Biotechnology
2009 Volume.37 No. 4 p.389 ~ p.398
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Development of pSJE6c, an Expression Vector for Kimchi Lactic Acid Bacteria, and Heterologous Gene Expression Using the Vector
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Lee Kang-Wook
Park Ji-Yeong
Lee Ji-Yeon
Lee Hwang-A
Baek Chang-Un
Jo Hyeon-Deok
Kim Joo-Yeon
Kwon Gun-Hee
Chun Ji-Yeon
Kim Jeong-Hwan
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Abstract
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Development of expression vectors is important for the basic and applied researches on kimchi LAB (lactic acid bacteria). An expression vector, pSJE6c was constructed by inserting P6C promoter sequence from Lactococcus lactis into pSJE, a shuttle vector for E. coli and Leuconostoc species. To test the efficiency of pSJE6c, aga (α-galactosidase) and lacZ (β-galactosidase) genes were expressed in Lactobacillus brevis 2.14. Compared to the pSJE, expression levels of both genes were increased, indicating P6C promoter was better than indigenous promoters. Enzyme activities of L. brevis cells harboring pSJE6caga (pSJE6c with aga) or pSJE6Z (pSJE6c with lacZ) were 1.5- 2 fold higher than those with pSJEaga (pSJE with aga) or pSJEZ (pSJE with lacZ). More RNA transcripts were detected in cells harboring pSJE6c based recombinant plasmid. The results indicated that heterologous gene expressions in kimchi LAB could be improved significantly by use of efficient expression vectors.
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KEYWORD
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Lactobacillus brevis, α-galactosidase, β-galactosidase, expression vectors, promoters
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