[巨大芽孢杆菌的芽孢-原生质体转化]

[Spore-spheroplast transformation of Bacillus megaterium].

作者信息

Takubo Y, Nishikawa J, Tani K, Nishihara T, Kondo M

机构信息

Faculty of Pharmaceutical Sciences, Osaka University.

出版信息

Nihon Saikingaku Zasshi. 1990 Sep;45(5):841-3. doi: 10.3412/jsb.45.841.

DOI:10.3412/jsb.45.841

PMID:2125086

Abstract

A new method for transformation of Bacillus megaterium was developed by modification of Chang and Cohen's method. In our method, spore spheroplasts were used as recipient cells instead of the protoplasts of vegetative cells. Longer incubation (60 min) of spore spheroplasts and plasmid DNA before treatment with polyethylene glycol remarkably increased the efficiency of transformation. The frequency of transformation was about 10(4) per microgram of plasmid DNA. A shot-gun-type cloning of chromosome DNA of B. megaterium ATCC 12872 was available in B. megaterium ATCC 19213 strain by this transformation method.

摘要

通过改进张和科恩的方法，开发出了一种用于巨大芽孢杆菌转化的新方法。在我们的方法中，使用芽孢原生质球作为受体细胞，而非营养细胞的原生质体。在用聚乙二醇处理之前，让芽孢原生质球与质粒DNA长时间孵育（60分钟），可显著提高转化效率。转化频率约为每微克质粒DNA 10⁴ 。通过这种转化方法，可在巨大芽孢杆菌ATCC 19213菌株中对巨大芽孢杆菌ATCC 12872的染色体DNA进行鸟枪法克隆。