假结核耶氏菌质粒DNA电穿孔转化试验

徐建国; 刘志奇; 俞东征

Abstract

徐建国,刘志奇,俞东征.假结核耶氏菌质粒DNA电穿孔转化试验[J].Chinese journal of Epidemiology,1990,11(4):239-242

假结核耶氏菌质粒DNA电穿孔转化试验

Plasmid DNA Transformation of Yersinia pseudotubercolosis by Electroporation

Received:December 27, 1989 Revised:January 20, 1990

DOI：

KeyWord: 电穿孔技术假结核耶氏菌鼠疫菌素

English Key Word: Electroporation Pesticin Y.pseudotubercolosis

FundProject:国家自然科学基金资助项目

Author Name	Affiliation
Xu Jianguo	Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing
刘志奇	Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing
俞东征	Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing

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Abstract:

本文首次报道了假结核耶氏菌的电穿孔转化技术。将编码鼠疫菌素的9.5kb的质粒克隆入pBR328的PstI位点,构建了质粒pJGX20。使用pJGX20,在电压为1300V、1400V,电容为25μF,电击时间为48~52微秒,使用10%甘油为电穿孔缓冲液,可获得每微克DNA 10⁸的转化子。pJGX20的鼠疫菌素基因,在假结核耶氏菌中得到良好表达。

English Abstract:

Rendering cells permeable to DNA molecules, electroporation involves the application of high intensity electric fields of short duration to reversibly change the biomembranes, which has been used successfully in some species of both gram negative and positive bacteria. We first reported the electro-tansformation technics of Yerstnia pseudotubercolosis. The plasmid pJGX20 was constiucted by cloning the 9.5 kb plasmid encording pesticin into PstI site of pBR328.Transformation efficiency of 10³ trans-formants/fig DNA was obtained at voltage of 1300V or 1400V,capicite of 25 μF,time constant of 48-52 msec, with 10% glycerol as electroporation buffer. The pesticin gene of pJGX20 was expressed well in Y. pseudotubercolosis.

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