酿酒酵母丙酮酸脱羧酶基因的克隆及表达Molecular cloning and expression of pyruvate decarboxylase cDNA from Saccharomyces cerevisiae
蒋雅红,游松,任杰,谢兰漪
JIANG Ya-hong 1; YOU Song 1; REN Jie 1; XIE Lan-yi 2 (1.School of Pharmaceutical Engineering;
摘要(Abstract)：
目的酿酒酵母 (Saccharomycescerevisiae)丙酮酸脱羧酶 (PDC)基因的克隆和表达。方法利用PCR技术从酿酒酵母的总cDNA中钓取出PDCsc基因 ,构建其高效表达质粒 ,利用Ion和ompT蛋白酶缺陷株EscherichiacoliBL2 1 (DE3)进行表达。结果扩增出长约 1 7kb的PDCsc基因 ,成功构建其表达质粒pSC -2 2b ,对转化菌株的SDS -PAGE分析结果显示 :该基因获得高效表达 ,表达蛋白占细胞总蛋白的 1 8 6 %。结论成功构建高效表达PDCsc基因的工程菌株 ,为实现利用PDCsc进行的生物转化奠定基础
Objective To clone and express the pyruvate decarboxylase(PDCsc) from Saccharomyces cerevisiae.Methods To clone PDCsc gene from its cDNA mixture by PCR, construct its expressing plasmid and express its encoding protein by using the Ion and ompT protease deficient strain Escherichia coli BL21(DE3). Results PDCsc gene (ca. 1.7 kb) was obtained,and its expressing plasmid pSC-22b was successfully constructed. The SDS-PAGE result showed that the overexpression of PDCsc was achieved and the target protein reached 18.6% among the total soluble proteins in its host strain.Conclusions The engineering strain for overexpression PDCsc was successfully constructed, which set up a base for the biotransformation using PDCsc as a catalyst.
关键词(KeyWords)：
酿酒酵母;丙酮酸脱羧酶;克隆;高效表达
Saccharomyces cerevisiae; pyruvate decarboxylase; clone; overexpression
基金项目(Foundation):
作者(Authors):
蒋雅红,游松,任杰,谢兰漪
JIANG Ya-hong 1; YOU Song 1; REN Jie 1; XIE Lan-yi 2 (1.School of Pharmaceutical Engineering;
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