一种质粒与枯草芽孢杆菌膜组分的特异性体外结合。
Specific in vitro binding of a plasmid to a membrane fraction of Bacillus subtilis.
作者信息
Korn R, Winston S, Tanaka T, Sueoka N
出版信息
Proc Natl Acad Sci U S A. 1983 Jan;80(2):574-8. doi: 10.1073/pnas.80.2.574.
A model system has been developed to study the in vitro binding of a plasmid to the membrane fraction from Bacillus subtilis. The plasmid DNA molecule used in these studies was pSL103 (8.0 kilobases), a chimeric plasmid consisting of a Staphylococcus aureus plasmid (pUB110, 4.5 kilobases) and a DNA fragment (3.5 kilobases) from Bacillus pumilus carrying trpC+ gene. This plasmid replicates in B. subtilis cells, and its in vivo membrane binding (as well as its replication) is dependent on the product of a DNA initiation gene, dna-1, of B. subtilis. In this paper we demonstrate the in vitro specific binding of exogenous pSL103 to the isolated membrane fraction. This in vitro binding is specific to the origin-containing portion (pUB110) of pSL103. The trpC+-carrying portion neither binds to the membrane fraction nor competes with pSL103 for binding to the membrane fraction in vitro. ColE1 plasmid, which does not replicate in B. subtilis, neither binds to the B. subtilis membrane fraction nor competes with pSL103 for binding.
已开发出一种模型系统，用于研究质粒在体外与枯草芽孢杆菌膜组分的结合。这些研究中使用的质粒DNA分子是pSL103（8.0千碱基），它是一种嵌合质粒，由金黄色葡萄球菌质粒（pUB110，4.5千碱基）和携带trpC+基因的短小芽孢杆菌的一个DNA片段（3.5千碱基）组成。该质粒在枯草芽孢杆菌细胞中复制，其体内膜结合（以及复制）依赖于枯草芽孢杆菌的一个DNA起始基因dna-1的产物。在本文中，我们证明了外源pSL103与分离的膜组分的体外特异性结合。这种体外结合对pSL103含起始点的部分（pUB110）具有特异性。携带trpC+的部分既不与膜组分结合，也不在体外与pSL103竞争结合膜组分。在枯草芽孢杆菌中不复制的ColE1质粒，既不与枯草芽孢杆菌膜组分结合，也不与pSL103竞争结合。
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