The Bacillus subtilis phoAIV gene: effects of in vitro inactivation on total alkaline phosphatase production.
The Bacillus subtilis phoAIV gene: effects of in vitro inactivation on total alkaline phosphatase production.
复制标题
枯草芽孢杆菌 phoAIV 基因：体外灭活对总碱性磷酸酶产生的影响。
DOI：
10.1016/0378-1119(90)90346-s
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发表时间：
1990
期刊：
影响因子：
3.5
通讯作者：
Hulett,FM
中科院分区：
文献类型：
--
作者：
Kapp,NV;Edwards,CW;Chesnut,RS;Hulett,FM
从成熟的碱性磷酸酶IV（APase IV）蛋白的前19个氨基酸推导出的退化寡脱氧核苷酸探针用于克隆枯草芽孢杆菌phoAIV基因编码区内部的DNA片段。用含有克隆DNA片段的整合质粒pJM 103在phoAIV基因座中构建插入突变。经Western-blot分析，phoAIV基因被阻断的菌株未检测到APase IV产物。阻断phoAIV对B.在磷酸盐饥饿和孢子形成培养条件下分析枯草杆菌168。突变inphoAIV减少了75%的磷酸盐饥饿的细胞中的总AP酶的特异性活性，并导致在消除盐可提取的膜AP酶，以及分泌的AP酶IV。对这种膜AP酶的分析表明，它是定位于裂解细胞的膜部分内而不分泌的aphoAIV基因产物。对产孢APase无影响。将ThephoAIV：：pJM 103插入作图并确定位于约1000 bp处。73°在B。360 °染色体。
A degenerative oligodeoxyribonucleotide probe deduced from the first 19 amino acids of the mature alkaline phosphatase IV (APase IV) protein was used to clone a DNA fragment internal to the coding region of thephoAIVgene ofBacillus subtilis. An insertional mutation was constructed in thephoAIVlocus using the integrative plasmid, pJM103, containing the cloned DNA fragment. The strain with the interruptedphoAIVgene showed no detectable APase IV product on Western-blot analysis. The impact of thephoAIVinterruption on total APase production inB. subtilis168 was analyzed under both phosphate starvation and sporulation culturing conditions. The mutation inphoAIVreduced total APase-specific activity by 75% in phosphate-starved cells, and resulted in the elimination of a salt-extractable membrane APase, as well as the secreted APase IV. Analysis of this membrane APase indicated that it is aphoAIVgene product which is localized within the membrane fraction of the lysed cell and not secreted. There was no effect on the production of sporulation APase. ThephoAIV::pJM103 insertion was mapped and determined to be located at approx. 73° on theB. subtilis360° chromosome.