INSERTION OF FOREIGN DNA INTO PLASMIDS FROM GRAM-POSITIVE BACTERIA INDUCES FORMATION OF HIGH-MOLECULAR-WEIGHT PLASMID MULTIMERS
INSERTION OF FOREIGN DNA INTO PLASMIDS FROM GRAM-POSITIVE BACTERIA INDUCES FORMATION OF HIGH-MOLECULAR-WEIGHT PLASMID MULTIMERS
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DOI：
10.1128/jb.170.3.1183-1190.1988
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发表时间：
1988-03-01
影响因子：
3.2
通讯作者：
EHRLICH, SD
中科院分区：
文献类型：
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作者：
GRUSS, A;EHRLICH, SD
质粒pUB 110、pC 194、pE 194和pT 181通常用作枯草芽孢杆菌和金黄色葡萄球菌中的克隆载体。我们报告说，外源DNA插入到这些质粒的任何一个结果在重组的高分子量质粒多聚体（HMW）的产生，目前作为串联头到尾拷贝。在野生型B中检测到HMW。subtilis和S.金黄色葡萄球菌HMW的产生取决于DNA插入的性质。大肠杆菌DNA的扩增，例如，pBR 322或pUC 18的插入片段产生大量HMW，而S.相同大小的金黄色葡萄球菌DNA对质粒谱没有影响。HMW的产生取决于质粒复制的模式;通过单链DNA中间体复制的质粒在外源DNA插入时产生HMW，而不产生单链DNA的质粒pAM β 1不产生HMW。我们认为HMW是滚环复制终止受损的产物，这种损伤是由于DNA插入的性质。
Plasmids pUB110, pC194, pE194, and pT181 are commonly used as cloning vectors in both Bacillus subtilis and Staphylococcus aureus. We report that insertion of foreign DNA into any of these plasmids results in the generation of high-molecular-weight plasmid multimers (HMW) of the recombinant, present as tandem head-to-tail copies. HMW was detected in wild-type B. subtilis and S. aureus strains. The production of HMW depended on the nature of the DNA insertion. Inserts of Escherichia coli DNA, e.g., pBR322 or pUC18, resulted in large amounts of HMW, whereas some inserts of S. aureus DNA of the same size had no effect on plasmid profile. The generation of HMW depended on the mode of plasmid replication; plasmids which replicate via a single-stranded DNA intermediate produced HMW upon foreign DNA insertion, whereas plasmid pAM.beta.1, which does not generate single-stranded DNA, did not generate HMW. We propose that HMW is a product of impaired termination of rolling-circle replication and that the impairment is due to the nature of the DNA insertion.