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构建一种用于香港海鸥型菌(一种与肠胃炎相关的新型细菌)的诱导型表达穿梭载体。

Construction of an inducible expression shuttle vector for Laribacter hongkongensis, a novel bacterium associated with gastroenteritis.

作者信息

Woo Patrick C Y, Ma Shirley S L, Teng Jade L L, Li Maria W S, Kao Richard Y T, Lau Susanna K P, Yuen Kwok-Yung

机构信息

Department of Microbiology, Faculty of Medicine, The University of Hong Kong, Queen Mary Hospital Compound, Pokfulam Road, Hong Kong.

出版信息

FEMS Microbiol Lett. 2005 Nov 1;252(1):57-65. doi: 10.1016/j.femsle.2005.08.026. Epub 2005 Sep 1.

Abstract

An Escherichia coli-Laribacter hongkongensis shuttle vector (pPW380) was constructed by ligating the 4701-bp EcoRI digested fragment of pHLHK8 to EcoRI digested pBK-CMV. An E. coli-L. hongkongensis inducible expression shuttle vector was further constructed by ligating a 2105-bp fragment that contains the tetracycline repressor and tetracycline-inducible promoter region of pALC2084 to the 8897-bp fragment of pPW380, deletion of the green fluorescent protein gene, and insertion of a multiple cloning site. This inducible expression system was able to express two commonly used reporter genes, the green fluorescent protein gene and the glutathione S-transferase gene, efficiently in E. coli and L. hongkongensis.

摘要

通过将pHLHK8经EcoRI酶切的4701 bp片段与经EcoRI酶切的pBK-CMV连接,构建了一种大肠杆菌-香港利比菌穿梭载体(pPW380)。通过将包含pALC2084的四环素阻遏物和四环素诱导型启动子区域的2105 bp片段连接到pPW380的8897 bp片段上,缺失绿色荧光蛋白基因,并插入多克隆位点,进一步构建了一种大肠杆菌-香港利比菌诱导型表达穿梭载体。该诱导型表达系统能够在大肠杆菌和香港利比菌中高效表达两种常用的报告基因,即绿色荧光蛋白基因和谷胱甘肽S-转移酶基因。

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