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[克隆到丝状M13mp8和M13WB2348噬菌体中的大肠杆菌rpo B基因的单向取向]

[Unidirectional orientation of the rpo B gene of E. coli cloned into filamentous M13mp8 and M13WB2348 phages].

作者信息

Paton E B, Vudmaska M I, Sverdlov E D

出版信息

Bioorg Khim. 1984 Nov;10(11):1544-7.

PMID:6098281
Abstract

E. coli DNA fragment containing the rpoB gene with an rpoB3 rifampicin resistance dominant mutation (coding for the beta-subunit of RNA polymerase), genes rpI J and rpI L coding for the ribosomal proteins L7/L12 and L10, and promoters determining transcription of all these genes were cloned in M13mp8 and WB2348 filamentous phages. E. coli cells containing recombinant phages acquired resistance to rifampicin up to its 600 micrograms/ml concentration. When cloned into M13mp8 and WB2348 phages, the given fragment is oriented in such a way that the direction of the transcription initiated from its own promoter coincides with that initiated from the lac UV5 promoter. In both cases the recombinant phages have no stable rifampicin resistance which is coded by the fragment.

摘要

含有rpoB基因(带有rpoB3利福平抗性显性突变,编码RNA聚合酶的β亚基)、编码核糖体蛋白L7/L12和L10的rpI J和rpI L基因以及决定所有这些基因转录的启动子的大肠杆菌DNA片段被克隆到M13mp8和WB2348丝状噬菌体中。含有重组噬菌体的大肠杆菌细胞对利福平的抗性可达其600微克/毫升浓度。当克隆到M13mp8和WB2348噬菌体中时,给定片段的取向使得从其自身启动子起始的转录方向与从lac UV5启动子起始的转录方向一致。在这两种情况下,重组噬菌体都没有该片段编码的稳定利福平抗性。

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