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Autoinduction of RpoS biosynthesis in the biocontrol strain Pseudomonas sp. M18.

机译:生防毒株假单胞菌(Pseudomonas sp。)中RpoS生物合成的自诱导。 M18。

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摘要

The rpoS gene from Pseudomonas sp. M18, which encodes predicted protein (an alternative sigma factor s, sigma S, or sigma 38) with 99.5% sequence identity with RpoS from Pseudomonas aeruginosa PAO1, was first cloned. In order to investigate the mechanism of rpoS expression, an rpoS null mutant, named M18S, was constructed with insertion of aacC1 cassette bearing a gentamycin resistance gene. With introduction of a plasmid containing an rpoS'-'lacZ translational fusion (pMERS) to wild-type strain M18 or M18S, it was first found that beta -galactosidase activity expressed in strain M18S (pMERS) decreased to fourfold of that expressed in the strain M18 (pMERS). When strain M18S (pMERS) was introduced with another plasmid pBBS containing the wild-type rpoS gene, its beta -galactosidase expression level was enhanced and almost restored to that in strain M18 (pMERS). Similarly, expression of beta -galactosidase from a chromosomal fusion of the promoter of the wild-type rpoS gene with lacZ (rpoS-lacZ) was enhanced fivefold in the presence of a plasmid with the wild-type rpoS gene. With these findings, it is suggested that RpoS sigma factor may be involved in autoinducing its own gene expression in Pseudomonas sp. M18..
机译:来自假单胞菌sp。的rpoS基因。首先克隆了M18,该蛋白编码与铜绿假单胞菌PAO1的RpoS具有99.5%序列同一性的预测蛋白(一个替代的sigma因子s,sigma S或sigma 38)。为了研究rpoS表达的机制,通过插入带有庆大霉素抗性基因的aacC1盒,构建了一个名为M18S的rpoS无效突变体。通过将含有rpoS'-'lacZ翻译融合(pMERS)的质粒引入野生型M18或M18S,首先发现在M18S(pMERS)菌株中表达的β-半乳糖苷酶活性降至在M18S(pMERS)中表达的β-半乳糖苷酶活性的四倍。菌株M18(pMERS)。当将菌株M18S(pMERS)与另一个包含野生型rpoS基因的质粒pBBS引入时,其β-半乳糖苷酶的表达水平得以提高,几乎恢复到菌株M18(pMERS)中。类似地,在具有野生型rpoS基因的质粒的存在下,来自野生型rpoS基因的启动子与lacZ(rpoS-lacZ)的染色体融合的β-半乳糖苷酶的表达提高了五倍。有了这些发现,建议RpoS sigma因子可能参与在Pseudomonas sp中自动诱导其自身的基因表达。 M18 ..

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