Mutoh N, Oosawa K, Simon M I
J Bacteriol. 1986 Sep;167(3):992-8. doi: 10.1128/jb.167.3.992-998.1986.
Hydroxylamine mutagenesis was used to alter the tar gene that encodes the transmembrane Tar protein required for chemotaxis. Mutants defective in chemotaxis were selected, and the mutation was characterized by DNA sequencing. Two classes of mutations were found: nonsense and missense. The nonsense mutations were distributed throughout the gene, while the missense mutations were found to cluster in a region that includes 185 amino acids at the C-terminal end of the Tar protein. Partial characterization of mutant phenotypes suggested that some are completely defective in signaling while responding to attractants and repellents by differential methylation. Other mutants are undermethylated and constantly tumble, while yet another class of mutants is overmethylated and biased toward constant swimming with little or no tumbling. These mutants will be useful in experiments designed to understand the mechanism of chemotaxis.
利用羟胺诱变来改变编码趋化作用所需跨膜Tar蛋白的tar基因。筛选出趋化作用有缺陷的突变体,并通过DNA测序对突变进行表征。发现了两类突变:无义突变和错义突变。无义突变分布在整个基因中,而错义突变则集中在Tar蛋白C末端包含185个氨基酸的区域。对突变体表型的部分表征表明,一些突变体在通过差异甲基化对引诱剂和驱避剂作出反应时信号传导完全有缺陷。其他突变体甲基化不足且持续翻滚,而另一类突变体甲基化过度且倾向于持续游动,很少或不翻滚。这些突变体将有助于开展旨在理解趋化作用机制的实验。