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枯草芽孢杆菌中的限制与修饰:枯草芽孢杆菌噬菌体SPR克隆甲基转移酶基因在大肠杆菌和枯草芽孢杆菌中的表达。

Restriction and modification in Bacillus subtilis: expression of the cloned methyltransferase gene from B. subtilis phage SPR in E. coli and B. subtilis.

作者信息

Montenegro M A, Pawlek B, Behrens B, Trautner T A

出版信息

Mol Gen Genet. 1983;189(1):17-20. doi: 10.1007/BF00326049.

Abstract

Expression of the SPR methyltransferase gene from B. subtilis phage SPR cloned into lambda and SPP1 was studied by analyzing the sensitivity of the hybrid phage DNAs to restriction by the enzymes HaeIII, MspI, and HpaII. The following results were obtained: (1) The genes were expressed both in the homologous (B. subtilis) and heterologous (E. coli) host. (2) The specificity of the expression of the cloned gene was identical to that of the gene in SPR. (3) Expression depended on the orientation of the cloned segment within the vector DNAs suggesting that vector promoters were involved in transcription. The coding strand of the cloned DNA was identified through hybridization with SPR mRNA.

摘要

通过分析杂交噬菌体DNA对HaeIII、MspI和HpaII酶切的敏感性,研究了枯草芽孢杆菌噬菌体SPR的SPR甲基转移酶基因克隆到λ噬菌体和SPP1噬菌体中的表达情况。得到以下结果:(1)这些基因在同源宿主(枯草芽孢杆菌)和异源宿主(大肠杆菌)中均有表达。(2)克隆基因的表达特异性与SPR中基因的表达特异性相同。(3)表达取决于克隆片段在载体DNA中的方向,这表明载体启动子参与了转录。通过与SPR mRNA杂交鉴定了克隆DNA的编码链。

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