Campanile C, Forlani G, Basso A L, Marasco R, Ricca E, Sacco M, Ferrara L, De Felice M
Istituto Adattamento Bovini e Bufali all'Ambiente del Mezzogiorno, Consiglio Nazionale delle Ricerche, Naples, Italy.
Appl Environ Microbiol. 1993 Feb;59(2):519-22. doi: 10.1128/aem.59.2.519-522.1993.
A genomic DNA library of the rumen bacterium Streptococcus bovis was constructed in Escherichia coli, and recombinant plasmids able to complement proA and proB mutations of the host were found. Southern hybridization and restriction analysis showed that a 3.5-kb fragment of S. bovis DNA contained two genes, organized in an operon and coding for enzymes functionally similar to the glutamyl phosphate reductase-glutamyl kinase enzyme complex that in E. coli catalyzes the first step of proline biosynthesis. Complementation of the E. coli mutations was observed with the fragment inserted in both orientations, which suggested that the S. bovis proBA operon was transcribed from its own promoter. Genetic and biochemical data suggested that the proline biosynthetic pathway of S. bovis is similar to the one previously characterized for E. coli.
构建了瘤胃细菌牛链球菌在大肠杆菌中的基因组DNA文库,并发现了能够互补宿主proA和proB突变的重组质粒。Southern杂交和限制性分析表明,牛链球菌DNA的一个3.5kb片段包含两个基因,它们以操纵子形式组织,编码的酶在功能上类似于大肠杆菌中催化脯氨酸生物合成第一步的谷氨酰磷酸还原酶-谷氨酰激酶酶复合物。当片段以两种方向插入时都观察到了对大肠杆菌突变的互补作用,这表明牛链球菌proBA操纵子是从其自身启动子转录的。遗传和生化数据表明,牛链球菌的脯氨酸生物合成途径与先前在大肠杆菌中表征的途径相似。