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[嗜中芽孢杆菌限制修饰基因在大肠杆菌中的克隆]

[Cloning of the restriction-modification genes of Bacillus centrosporus in Escherichia coli].

作者信息

Povilenis P I, Lubis A A, Ianulaĭtis A A

出版信息

Genetika. 1988 Feb;24(2):210-5.

PMID:2834268
Abstract

A genomic library of Bacillus centrosporus was obtained using pBR327 as a vector. The total plasmid DNA of the library was cleaved by the BcnI restriction endonuclease and then transformed in Escherichia coli RR1. Two clones possessing restriction and DNA modification profiles of BcnI were identified among the transformants. Their respective plasmids were 13.3 and 9.05 kbp in size. Restriction mapping of both plasmids showed each of them to contain two sites for HindIII and one for both Eco31I and Eco47III, located at the same distance. This was assumed to be the location region of the BcnI restriction-modification genes. Confirmation of the assumption was obtained by deletion mapping of the recombinant plasmids. Special features concerning cloning of the restriction-modification genes are discussed on the basis of the results obtained.

摘要

以pBR327为载体构建了芽孢杆菌的基因组文库。该文库的总质粒DNA用BcnI限制性内切酶切割,然后转化到大肠杆菌RR1中。在转化子中鉴定出两个具有BcnI限制性和DNA修饰图谱的克隆。它们各自的质粒大小分别为13.3和9.05kbp。两种质粒的限制性图谱显示,它们各自含有两个HindIII位点以及一个Eco31I和Eco47III位点,且位置相同。这被认为是BcnI限制性修饰基因的定位区域。通过重组质粒的缺失图谱分析证实了这一假设。根据所得结果讨论了限制性修饰基因克隆的特殊特征。

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