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通过诱导克隆的φX174基因来裂解大肠杆菌。

Lysis of Escherichia coli by induction of cloned phi X174 genes.

作者信息

Henrich B, Lubitz W, Plapp R

出版信息

Mol Gen Genet. 1982;185(3):493-7. doi: 10.1007/BF00334146.

Abstract

The largest of the fragments produced by AluI digestion of phi X174 RFI DNA comprises genes E and J as well as parts of genes D and F. This DNA fragment (1007 bp) was cloned into the lac z' gene of plasmid pUR222. In the recombinant plasmid pUH12, transcription of the phi X174 genes is controlled by the lac p-o region. Induction of the cloned genes by addition of the lac inducer, IPTG, resulted in lysis of the bacteria. Cloning of the corresponding AluI-fragment from phi X174am3 DNA, carrying an amber mutation in gene E, showed that the expression of this gene alone is sufficient to trigger cell lysis. The time interval between the addition of IPTG and the onset of lysis depended on the concentration of the inducer, however, the rate of lysis was similar at all IPTG concentrations used.

摘要

用AluI消化φX174 RFI DNA产生的最大片段包含基因E和J以及基因D和F的部分。这个DNA片段(1007 bp)被克隆到质粒pUR222的lac z'基因中。在重组质粒pUH12中,φX174基因的转录由lac p-o区域控制。通过添加lac诱导剂IPTG诱导克隆基因会导致细菌裂解。从携带基因E琥珀突变的φX174am3 DNA中克隆相应的AluI片段,结果表明仅该基因的表达就足以引发细胞裂解。添加IPTG到裂解开始的时间间隔取决于诱导剂的浓度,然而,在所有使用的IPTG浓度下裂解速率相似。

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