PLASMID DELETION FORMATION BETWEEN SHORT DIRECT REPEATS IN BACILLUS-SUBTILIS IS STIMULATED BY SINGLE-STRANDED ROLLING-CIRCLE REPLICATION INTERMEDIATES
PLASMID DELETION FORMATION BETWEEN SHORT DIRECT REPEATS IN BACILLUS-SUBTILIS IS STIMULATED BY SINGLE-STRANDED ROLLING-CIRCLE REPLICATION INTERMEDIATES
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DOI：
10.1007/bf00273591
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发表时间：
1991-04-01
期刊：
影响因子：
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通讯作者：
PEETERS, BPH
中科院分区：
文献类型：
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作者：
BRON, S;HOLSAPPEL, S;PEETERS, BPH
研究了滚环复制模式和单链 DNA (ss DNA) 的产生对枯草芽孢杆菌中短同向重复序列之间质粒缺失形成的影响。由直接重复序列（9、18 或 27 bp）组成的缺失单元（侧翼有或没有反向重复序列（300 bp））被引入各种质粒复制子中，产生不同量的单链 DNA（总质粒 DNA 的 0% 到 40%）。对于产生单链DNA的滚环型质粒，同向重复之间的缺失频率比不产生单链DNA的质粒高3至13倍。当正向重复序列侧翼为反向重复序列时，ss DNA 生成质粒中的缺失频率增加了 20 至 140 倍。这些结果支持基于滚环型质粒互补链合成过程中模板转换错误的缺失形成模型。在存在互补链合成的功能性起始位点（负起点）的情况下，枯草芽孢杆菌中的 ss DNA 生成质粒 pTA1060 的结构不稳定性（短同向重复之间的缺失形成）非常低。这一观察结果表明，开发枯草芽孢杆菌稳定的宿主载体克隆系统是可能的。
The effects of the rolling-circle mode of replication and the generation of single-stranded DNA (ss DNA) on plasmid deletion formation between short direct repeats in Bacillus subtilis were studied. Deletion units consisting of direct repeats (9, 18, or 27 bp) that do or do not flank inverted repeats (300 bp) were introduced into various plasmid replicons that generate different amounts of ss DNA (from 0% to 40% of the total plasmid DNA). With ss DNA-generating rolling-circle-type plasmids, deletion frequencies between the direct repeats were 3- to 13-fold higher than in plasmids not generating ss DNA. When the direct repeats flanked inverted repeats the deletion frequencies in ss DNA-generating plasmids were increased by as much as 20- to 140-fold. These results support models for deletion formation based on template-switching errors during complementary strand synthesis of rolling-circle-type plasmids. The structural instability (deletion formation between short direct repeats) of the ss DNA-generating plasmid pTA1060 in B. subtilis was very low in the presence of a functional initiation site for complementary strand synthesis (minus origin). This observation suggests that it will be possible to develop stable host-vector cloning systems for B. subtilis.