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人谷胱甘肽S-转移酶和铜/锌超氧化物歧化酶在乳酸乳球菌中的食品级表达

Food-grade expression of human glutathione S-transferase and Cu/Zn superoxide dismutase in Lactococcus lactis.

作者信息

Xiang Hua, Wei Wenzhong, Tan Huarong

机构信息

Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.

出版信息

Biomol Eng. 2003 Mar;20(3):107-12. doi: 10.1016/s1389-0344(03)00007-8.

Abstract

A food-grade gene expression system in Lactococcus lactis was established by the combination of a vector containing the lacF gene as the selection marker and a strain WZ103 carrying an in-frame deletion of this gene in the chromosome as the host. The human glutathione S-transferase A1-1 (hGSTA1) and Cu/Zn superoxide dismutase (hSOD) genes were respectively cloned into a food-grade vector under the control of the lactococcal inducible promoter P(lacA). The resulting expression plasmids were separately introduced into the lactose-deficient (Lac(-)) host, and the lactose-utilizing (Lac(+)) transformants were directly selected on a chemically defined medium, using lactose as the sole carbon source. The successful food-grade expression of hGSTA1 and hSOD in the L. lactis WZ103 transformed with these plasmids were analyzed by Western blotting and enzymatic activity assay, respectively.

摘要

通过将含有lacF基因作为选择标记的载体与一株在染色体上该基因发生框内缺失的菌株WZ103作为宿主相结合,在乳酸乳球菌中建立了一种食品级基因表达系统。将人谷胱甘肽S-转移酶A1-1(hGSTA1)和铜/锌超氧化物歧化酶(hSOD)基因分别克隆到在乳酸乳球菌诱导型启动子P(lacA)控制下的食品级载体中。将所得表达质粒分别导入乳糖缺陷型(Lac(-))宿主中,并在化学成分确定的培养基上直接选择利用乳糖的(Lac(+))转化体,使用乳糖作为唯一碳源。分别通过蛋白质免疫印迹和酶活性测定分析了用这些质粒转化的乳酸乳球菌WZ103中hGSTA1和hSOD的成功食品级表达。

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