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枯草芽孢杆菌中的转化:DNA结合缺陷型突变体的特性

Transformation in Bacillus subtilis: properties of DNA-binding-deficient mutants.

作者信息

Smith H, de Vos W, Bron S

出版信息

J Bacteriol. 1983 Jan;153(1):12-20. doi: 10.1128/jb.153.1.12-20.1983.

Abstract

Transformation-deficient mutants of Bacillus subtilis were selected after replica plating on agar plates containing transforming DNA. Out of 24 mutants tested, 3 showed highly reduced abilities to bind donor DNA; the residual levels of binding were similar to those of noncompetent cells. Transformation and transfection were reduced to nondetectable levels in the mutants. However, transduction with phage SPP1 occurred at normal frequencies. The nuclease activities involved in entry of donor DNA were present in the mutants. Comparison of protein patterns by two-dimensional gel electrophoresis revealed the absence of one major protein in the mutants as compared with the wild-type strain. This protein (molecular weight, approximately 18,000; isoelectric point, 5.0) appeared to be membrane associated. The protein was specific for competent cells, suggesting that it is involved in the binding of donor DNA.

摘要

在含有转化DNA的琼脂平板上进行影印接种后,筛选出了枯草芽孢杆菌的转化缺陷型突变体。在测试的24个突变体中,有3个显示出结合供体DNA的能力大幅降低;其残留的结合水平与非感受态细胞相似。这些突变体中的转化和转染降低到了无法检测的水平。然而,用噬菌体SPP1进行转导的频率正常。突变体中存在参与供体DNA进入的核酸酶活性。通过二维凝胶电泳对蛋白质模式进行比较,发现与野生型菌株相比,突变体中缺少一种主要蛋白质。这种蛋白质(分子量约为18,000;等电点为5.0)似乎与膜相关。该蛋白质对感受态细胞具有特异性,表明它参与供体DNA的结合。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/823a/217336/c7188582eb4c/jbacter00248-0037-a.jpg

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