移动单孢菌丙酮酸脱羧酶基因的克隆及表达

目的克隆与表达移动单孢菌(Zymomonas mobilis)丙酮酸脱羧酶(Pyruvate decarboxylase,PDC)基因。方法利用聚合酶链反应(PCR)技术从移动单孢的基因组DNA中扩增出PDCzm基因,构建其高效表达质粒,利用Ion和ompT蛋白酶缺陷株Escherichia coli BL21(DE3)进行表达。结果扩增出DNA碱基对数目约1.7 kb的PDCzm基因,成功构建其表达质粒pZM 22 b,对转化菌株的变性聚丙烯酰胺凝胶电泳(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37.9%。结论成功构建高效表达PDCzm基因的工程菌株,为实现利用PDCzm进行的生物转化奠定了基础。

Molecular cloning and expression of pyruvate decarboxylase gene from Zymomonas mobilis

Objective To clone and express the pyruvate decarboxylase(PDCzm) from Zymomonas mobilis.Methods To clone PDCzm gene from its genomic DNA by PCR,construct its expressing plasmid and express its encoding protein by using Ion and ompT protease deficient strain Escherichia coli BL21(DE3).Results PDCzm gene(ca.1.7kb) was obtained and its expressing plasmid pZM-22b was successfully constructed.SDS-PAGE result showed that the overexpression of PDCzm was achieved and the target protein reached 37.9% among the total proteins in its host strain.Conclusions The engineering strain for overexpression PDCzm was successfully constructed,which set up a base for the biotransformation using PDCzm as catalyst.