Environmental sensing in bacteria often involves the concerted action of sensor kinases and response regulators. Degenerate oligonucleotide primers were designed on the basis of amino acid similarity in the response regulators of these two-component systems. The primers were used in PCR to specifically amplify an internal DNA segment corresponding to the receiver module domain from genes encoding response regulators. Amplification products of the expected size were obtained from 12 different Gram-positive and Gram-negative bacteria. Sequence analysis revealed that 22 DNA fragments, which clearly originated from response regulator genes, were amplified from Escherichia coli, Agrobacterium tumefaciens, Bacillus subtilis and Lactobacillus bulgaricus. In each of these four species the receiver module of putative response regulator genes, which do not seem to be related to any of the already characterized genes, was identified. This simple and powerful method is therefore particularly useful for discovering new signal transduction systems which cannot be revealed by usual genetic studies.

译文

细菌中的环境传感通常涉及传感器激酶和响应调节剂的协同作用。简并寡核苷酸引物是根据这些两组分系统的响应调节剂中的氨基酸相似性设计的。引物用于PCR,以从编码反应调节剂的基因中特异性扩增与接收器模块结构域相对应的内部DNA片段。从12种不同的革兰氏阳性和革兰氏阴性细菌中获得了预期大小的扩增产物。序列分析表明,从大肠杆菌,根癌农杆菌,枯草芽孢杆菌和保加利亚乳杆菌中扩增出22个明显源自响应调节基因的DNA片段。在这四个物种中的每一个物种中,都鉴定了假定的反应调节基因的接收器模块,该模块似乎与任何已经表征的基因无关。因此,这种简单而强大的方法对于发现通常的遗传研究无法揭示的新信号转导系统特别有用。

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