Munson M A, Baumann L, Baumann P
Microbiology Section, University of California, Davis 95616-8665.
Gene. 1993 Dec 31;137(2):171-8. doi: 10.1016/0378-1119(93)90003-l.
The aphid Schizaphis graminum is dependent on an association with Buchnera aphidicola, an eubacterial endosymbiont located in specialized host cells. Past studies have indicated that Escherichia coli is the closest known relative of the endosymbiont which has many genetic attributes of free-living bacteria. In order to obtain information on the properties of highly expressed genes, we have chosen for study the single-copy rrs (gene encoding 16S rRNA) of B. aphidicola. A 4.4-kb DNA fragment was cloned into E. coli and the nucleotide (nt) sequence determined. Several ORFs were identified; the order of genes was argS-rrs-ORF1-rnh-dnaQ. ArgS, RNase H and DnaQ had 36-57% amino acid (aa) identity to the homologous proteins of E. coli. B. aphidicola rrs appears to be part of an operon consisting of a putative promoter, rrs and two inverted repeats resembling Rho-independent terminators. Comparisons of the sequences of argS-rrn DNA fragments from endosymbionts of six additional aphid species indicated conservation of sequences corresponding to a single -35 (TTGACA) and -10 (TGTAAT) promoter region, as well as boxA (sequence involved in antitermination) and boxC. The B. aphidicola argS-rrn DNA fragments from endosymbionts from seven species of aphids had promoter activities in E. coli which ranged from 6 to 135% of that observed with a comparable DNA fragment of E. coli rrnB. Similarly, the putative B. aphidicola terminator was functional in E. coli. In most eubacteria, the rRNA-encoding genes are arranged in the order, 16S, 23S, 5S, and are part of a single operon.(ABSTRACT TRUNCATED AT 250 WORDS)
麦二叉蚜依赖于与布赫纳氏菌的共生关系,布赫纳氏菌是一种位于特殊宿主细胞中的真细菌内共生体。过去的研究表明,大肠杆菌是这种内共生体已知的亲缘关系最近的物种,它具有许多自由生活细菌的遗传特性。为了获取高表达基因特性的信息,我们选择研究布赫纳氏菌的单拷贝rrs(编码16S rRNA的基因)。将一个4.4kb的DNA片段克隆到大肠杆菌中并测定核苷酸(nt)序列。鉴定出了几个开放阅读框(ORF);基因顺序为argS - rrs - ORF1 - rnh - dnaQ。精氨酸琥珀酰转移酶(ArgS)、核糖核酸酶H(RNase H)和DNA聚合酶III的ε亚基(DnaQ)与大肠杆菌的同源蛋白有36 - 57%的氨基酸(aa)同一性。布赫纳氏菌的rrs似乎是一个操纵子的一部分,该操纵子由一个假定的启动子、rrs和两个类似于不依赖于ρ因子的终止子的反向重复序列组成。对来自另外六种蚜虫内共生体的argS - rrn DNA片段序列的比较表明,对应于单个 - 35(TTGACA)和 - 10(TGTAAT)启动子区域以及boxA(抗终止相关序列)和boxC的序列是保守的。七种蚜虫内共生体的布赫纳氏菌argS - rrn DNA片段在大肠杆菌中具有启动子活性,其活性范围为大肠杆菌rrnB可比DNA片段所观察到活性的6%至135%。同样,假定的布赫纳氏菌终止子在大肠杆菌中是有功能的。在大多数真细菌中,编码rRNA的基因按16S、23S、5S的顺序排列,并且是单个操纵子的一部分。(摘要截短于250字)