Analysis of spore photoproduct lyase operon (splAB) function using targeted deletion-insertion mutations spanning the Bacillus subtilis operons ptsHI and splAB.
Analysis of spore photoproduct lyase operon (splAB) function using targeted deletion-insertion mutations spanning the Bacillus subtilis operons ptsHI and splAB.
复制标题
使用跨越枯草芽孢杆菌操纵子 ptsHI 和 splAB 的靶向删除-插入突变分析孢子光产物裂解酶操纵子 (splAB) 功能。
DOI：
10.1007/s004380050532
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发表时间：
1997
期刊：
影响因子：
--
通讯作者：
Fajardo-Cavazos,P
中科院分区：
文献类型：
--
作者：
Nicholson,WL;Chooback,L;Fajardo-Cavazos,P
发芽的枯草芽孢杆菌孢子部分利用孢子光产物裂解酶修复紫外线诱导的DNA损伤。SP裂解酶由splaboperon的第二个顺子ysplb编码。plaboperon在产孢过程中由P1启动子转录，P1启动子部分重叠于上游p2操作子的转录终止子，P1启动子反过来编码Hpr蛋白和PEP：糖磷酸转移酶（PTS）系统的酶I组分。为了确定这些连续操纵子的物理和功能边界，通过体外位点定向诱变在该区域产生零突变，其中克隆的ptsi - splab区域的部分被移除，取而代之的是抗生素抗性盒，然后通过转化引入b。细小。一个跨越ptsi、splA和splbabr的删除插入破坏了产生的突变体利用PTS糖葡萄糖的能力。去除splabone或splaand splb基因的缺失不影响葡萄糖的利用，这表明splabgene产物与PTS功能无关。利用缺失splab的缺失插入突变体建立了一个互补系统，该系统允许将克隆的splaboperon的等位基因放置在染色体上。利用互补体系探讨SP裂解酶在测定孢子抗紫外线能力中的作用。
GerminatingBacillus subtilisspores repair UV-induced DNA damage in part using the enzyme spore photoproduct (SP) lyase. SP lyase is encoded bysplB, the second cistron of thesplABoperon. ThesplABoperon is transcribed during sporulation from the P1 promoter, which partially overlaps the transcriptional terminator of the upstreamptsHIoperon, which in turn encodes the Hpr protein and Enzyme I components of the PEP:sugar phosphotransferase (PTS) system. In order to determine the physical and functional boundaries of these contiguous operons, null mutations were generated in the region by in vitro site-directed mutagenesis, in which parts of the clonedptsI-splABregion were removed and replaced with anermCantibiotic resistance cassette, then introduced by transformation intoB. subtilis. A deletion-insertion spanningptsI,splA, andsplBabolished the ability of the resulting mutant to utilize the PTS sugar glucose. Deletions removing eithersplBalone or bothsplAandsplBdid not affect glucose utilization, thus indicating thatsplABgene products are not involved in PTS function. A complementation system was developed using the deletion-insertion mutant lackingsplABwhich allows placement of alleles of the clonedsplABoperon at the chromsomalamyElocus. The complementation system was used to explore the role of SP lyase in determining spore UV resistance.