Hsu L M, Klee H J, Zagorski J, Fournier M J
J Bacteriol. 1984 Jun;158(3):934-42. doi: 10.1128/jb.158.3.934-942.1984.
A plasmid containing a gene for the most abundant Escherichia coli leucine isoacceptor tRNA, tRNALeu1 (anticodon CAG) was isolated from the Clarke-Carbon bank of cloned E. coli DNA. The clone contains a 12.3-kilobase DNA insert which was mapped by F' DNA hybridization analysis to the region 82 to 89 min on the chromosome. The cloned tDNALeu corresponds to the minor of two chromosomal regions containing different amounts of DNA complementary to tRNALeuCAG . Sequencing of the tDNA region revealed it to contain a multimeric transcription unit consisting of four different tRNA genes. The genes are in the arrangement 5'-leader- tRNAArgCCG -57 base pairs- tRNAHisGUG -20 base pairs- tRNALeuCAG -42 base pairs- tRNAProUGG -3'. Coordinate expression of the component tRNAs in vivo and the absence of intercistronic promoters indicated that all four tDNAs reside in the same operon. The tDNA sequence is bounded by a promoter element showing good agreement with the procaryotic consensus sequence and a GC-rich stem-loop element that corresponds to a rho-independent terminator. The promoter region contains a GC-rich sequence that agrees with a suggested consensus stringency control element and two domains possessing dyad symmetry which flank the Pribnow box and include the putative stringency control region.
从克隆的大肠杆菌DNA的克拉克-卡本文库中分离出一个含有编码大肠杆菌中最丰富的亮氨酸同工受体tRNA(tRNALeu1,反密码子CAG)基因的质粒。该克隆包含一个12.3千碱基的DNA插入片段,通过F' DNA杂交分析将其定位到染色体上82至89分钟的区域。克隆的tDNALeu对应于两个染色体区域中较小的那个,这两个区域含有不同量的与tRNALeuCAG互补的DNA。对tDNA区域进行测序发现它包含一个由四个不同tRNA基因组成的多聚体转录单位。这些基因的排列顺序为5'-前导序列-tRNAArgCCG-57个碱基对-tRNAHisGUG-20个碱基对-tRNALeuCAG-42个碱基对-tRNAProUGG-3'。体内各组成tRNA的协同表达以及不存在顺反子间启动子表明所有四个tDNA位于同一个操纵子中。tDNA序列由一个与原核生物共有序列高度一致的启动子元件和一个富含GC的茎环元件界定,该茎环元件对应于一个不依赖ρ因子的终止子。启动子区域包含一个与建议的共有严格控制元件一致的富含GC的序列,以及两个具有二重对称的结构域,它们位于普里布诺框两侧,包括假定的严格控制区域。