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[通过电穿孔法将质粒DNA转化到假结核耶尔森氏菌中]

[Plasmid DNA transformation of Yersinia pseudotuberculosis by electroporation].

作者信息

Xu J G

机构信息

Institute of Epidemiology and Microbiology, Chinese Academy of Preventive Medicine, Beijing.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 1990 Aug;11(4):239-42.

PMID:2225008
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-transformation technics of Yersinia pseudotuberculosis. The plasmid pJGX20 was constructed by cloning the 9.5 kb plasmid encoding pesticin into Pst I site of pBR328. Transformation efficiency of 103 transformants/micrograms DNA was obtained at voltage of 1300V or 1400V, capacity of 25 microF, time constant of 48-52 msec, with 10% glycerol as electroporation buffer. The pesticin gene of pJGX20 was expressed well in Y. pseudotuberculosis.

摘要

电穿孔法可使细胞对DNA分子具有通透性,该方法通过施加短时间的高强度电场来可逆地改变生物膜,已在一些革兰氏阴性菌和阳性菌中成功应用。我们首次报道了假结核耶尔森菌的电转化技术。质粒pJGX20是通过将编码杀有害素的9.5 kb质粒克隆到pBR328的Pst I位点构建而成。在电压为1300V或1400V、电容为25微法、时间常数为48 - 52毫秒、以10%甘油作为电穿孔缓冲液的条件下,获得了每微克DNA 103个转化子的转化效率。pJGX20的杀有害素基因在假结核耶尔森菌中表达良好。

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[Plasmids of Yersinia pseudotuberculosis].
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