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[将φ80噬菌体整合到异常染色体位点并分离出一种特异性转导噬菌体]

[Incorporation of phi80 phage into unusual chromosome sites and isolation of a specialized transducing bacteriophage].

作者信息

Il'ina T S, Nechaeva E V

出版信息

Genetika. 1977;13(12):2181-8.

PMID:355053
Abstract

The mutant strain KS713 of Escherichia coli K-12 deleted for the normal insertion site and secondary preferable one was obtained. The insertion frequency of phage phi80 into the double deletion strain is reduced about 30-fold with respect to integration into the strain H47 with deletion of the primary phi80 attachment site and about 500-fold relative to integration into wild type Escherichia coli. Analysis of the rare abnormal lysogens of KS 713 strain indicates that there are secondary sites on the chromosome, which are utilized for prophage attachment if insertion at preferable secondary att80-II site is eliminated too. The insertion of phi80 phage into the bfe locus was obtained by the appropriate selection technique. Induced prophage excision from the bfe site was rather efficient and lysates contained phi80 phage particles that could specificically transduce the argH+ gene. Upon transduction into a recipient strain carrying recA, heterogenotes harbouring both the wild-type and the mutant argH genes were isolated. These heterogenotes were used for producing high-frequency transducing lysates.

摘要

获得了大肠杆菌K-12的突变菌株KS713,该菌株缺失了正常插入位点和次要的优先插入位点。相对于整合到缺失主要phi80附着位点的H47菌株中,噬菌体phi80插入双缺失菌株的频率降低了约30倍,相对于整合到野生型大肠杆菌中降低了约500倍。对KS 713菌株罕见的异常溶原菌的分析表明,染色体上存在次要位点,如果在优先的次要att80-II位点的插入也被消除,这些位点将用于原噬菌体附着。通过适当的选择技术,获得了phi80噬菌体插入bfe基因座的情况。从bfe位点诱导原噬菌体切除相当有效,裂解物中含有能够特异性转导argH+基因的phi80噬菌体颗粒。转导到携带recA的受体菌株后,分离出了同时含有野生型和突变型argH基因的杂合子。这些杂合子被用于制备高频转导裂解物。

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