Silverman M, Simon M
J Bacteriol. 1977 Jun;130(3):1317-25. doi: 10.1128/jb.130.3.1317-1325.1977.
Molecular cloning techniques were used to construct Escherichia coli-lambda hybrids that contained many of the genes necessary for flagellar rotation and chemotaxis. The properties of specific hybrids that carried the classical "cheA" and "cheB" loci were examined by genetic complementation and by measuring the capacity of the hybrids to direct the synthesis of specific polypeptides. The results of these tests with lambda hybrids and with a series of deletion mutations derived from the hybrids redefined the "cheA" and "cheB" regions. Six genes were resolved: cheA, cheW, cheX, cheB, cheY, and cheZ. They directed the synthesis of specific polypeptides with the following apparent molecular weights: cheA, 76,000 and 66,000; cheW, 12,000; cheX, 28,000; cheB, 38,000; cheY, 8,000; and cheZ, 24,000. The presence of another gene, cheM, was inferred from the protein synthesis experiments. The cheM gene directed the synthesis of polypeptides with apparent molecular weights of 63,000, 61,000, and 60,000. The synthesis of all of these polypeptides is regulated by the same mechanisms that regulate the synthesis of flagellar-related structural components.
分子克隆技术被用于构建大肠杆菌-λ杂种,这些杂种包含鞭毛旋转和趋化作用所需的许多基因。通过遗传互补以及测量杂种指导特定多肽合成的能力,对携带经典“cheA”和“cheB”位点的特定杂种的特性进行了研究。用λ杂种以及源自杂种的一系列缺失突变体进行的这些测试结果,重新定义了“cheA”和“cheB”区域。分辨出了六个基因:cheA、cheW、cheX、cheB、cheY和cheZ。它们指导合成具有以下表观分子量的特定多肽:cheA,76,000和66,000;cheW,12,000;cheX,28,000;cheB,38,000;cheY,8,000;cheZ,24,000。从蛋白质合成实验推断出另一个基因cheM的存在。cheM基因指导合成表观分子量为63,000、61,000和60,000的多肽。所有这些多肽的合成均受调控鞭毛相关结构成分合成的相同机制的调节。