Todorova R T
Mol Biol (Mosk). 1993 Mar-Apr;27(2):429-35.
Three mutant forms of the ribosomal protein L7/L12 with Ser1, Met14, and Met26 replacements with Tyr were constructed for studying the N-terminal domain of the protein. Three point mutations in the gene L7/L12 on the phage M13mp18 were generated. Recombinant plasmids containing the mutant genes were constructed on the base of expression vector pKK223-3 and plasmid pUC19. The mutant proteins were expressed in Escherichia coli cells, and methods of their purification were developed. It was found that the mutant proteins L7/L12 do not differ from the wild-type protein L7/L12 and represent a suitable object for 1H-NMR study of the L7/L12 protein. The three mutant proteins bind with the E. coli ribosome.
构建了核糖体蛋白L7/L12的三种突变形式,其中Ser1、Met14和Met26被Tyr取代,用于研究该蛋白的N端结构域。在噬菌体M13mp18的L7/L12基因上产生了三个点突变。基于表达载体pKK223-3和质粒pUC19构建了含有突变基因的重组质粒。突变蛋白在大肠杆菌细胞中表达,并开发了其纯化方法。发现突变蛋白L7/L12与野生型蛋白L7/L12没有差异,是用于L7/L12蛋白1H-NMR研究的合适对象。这三种突变蛋白与大肠杆菌核糖体结合。