Zsebo K M, Hearst J E
Cell. 1984 Jul;37(3):937-47. doi: 10.1016/0092-8674(84)90428-8.
Transposon mutagenesis and complementation analysis of the photosynthesis genes in Rhodopseudomonas capsulata is presented utilizing Tn5.7 mutagenized R-primes. The R-prime pRPS404 contains many of the genes necessary for the differentiation of the photosynthetic apparatus. Utilizing homologous recombination, 30 independent copies of Tn5.7 were inserted into the R. capsulata chromosome with subsequent deletion of wild-type alleles. Mutants were characterized by absorption spectroscopy, SDS-PAGE, and determination of capability for photosynthetic growth. Many mutations in the bacteriochlorophyll and carotenoid biosynthetic pathways were isolated. A regulatory mutation was isolated affecting reaction-center synthesis as well as a 44 kd heme-containing polypeptide. Complementation analysis using various pRPR404::Tn5.7 plasmids has led to the postulation of transcriptional units.
本文利用Tn5.7诱变的R-质粒，对荚膜红假单胞菌光合作用基因进行转座子诱变和互补分析。R-质粒pRPS404包含光合装置分化所需的许多基因。利用同源重组，将30个独立的Tn5.7拷贝插入荚膜红假单胞菌染色体，随后野生型等位基因缺失。通过吸收光谱、SDS-PAGE和光合生长能力测定对突变体进行了表征。分离出许多细菌叶绿素和类胡萝卜素生物合成途径中的突变。分离出一个影响反应中心合成以及一种含44kd血红素多肽的调控突变。使用各种pRPR404::Tn5.7质粒进行的互补分析导致了转录单元的推测。
