Nakajima N, Ozeki H, Shimura Y
Cell. 1981 Jan;23(1):239-49. doi: 10.1016/0092-8674(81)90288-9.
The structure and organization on the Escherichia coli chromosome of the gene cluster coding for two methionine tRNAs (tRNAmMet), four glutamine tRNAs (two tRNA1Gln and two tRNA2Gln), and a previously unidentified tRNA (called tRNAx) have been studied by restriction enzyme analysis and DNA sequencing, utilizing a specialized transducing bacteriophage (lambda psu degrees 2) carrying the supB-supE region. From the sequence analysis, the previously unidentified tRNA has been shown to have an anticodon sequence (5'-UAG-3') corresponding to a leucine codon. The organization of this tRNA gene cluster on the E. coli chromosome is tRNAmMet-9 base pairs-tRNAx-23 base pairs-tRNA1Gln-34 base pairs-tRNA1Gln-15 base pairs-tRNAmMet-47 base pairs-tRNA2Gln-37 base pairs-tRNA2Gln. The duplicated genes coding for tRNAmMet, tRNA,Gln, and tRNA2Gln have identical sequences, which are the same as the sequences determined previously with tRNA molecules. These tRNA sequences are preceded by a single promoter region where a "Pribnow box" sequence is present seven base pairs upstream from the transcription start site. The spacer regions separating the seven tRNA sequences are different from each other both in size and in nucleotide sequence. The possible implication of these sequences for precursor processing is discussed. A restriction fragment that has been originally identified in lambda psu degrees 2 DNA and shown to contain the seven tRNA genes has been detected in the E. coli chromosome, thereby suggesting that this tRNA gene cluster is present in the bacterial genome with the same organization as in the transducing phage genome.
利用携带supB - supE区域的特异性转导噬菌体(λpsu°2),通过限制酶分析和DNA测序,对编码两种甲硫氨酸tRNA(tRNAmMet)、四种谷氨酰胺tRNA(两种tRNA1Gln和两种tRNA2Gln)以及一种先前未鉴定的tRNA(称为tRNAx)的基因簇在大肠杆菌染色体上的结构和组织进行了研究。通过序列分析,已表明先前未鉴定的tRNA具有与亮氨酸密码子相对应的反密码子序列(5'-UAG-3')。该tRNA基因簇在大肠杆菌染色体上的组织形式为:tRNAmMet - 9个碱基对 - tRNAx - 23个碱基对 - tRNA1Gln - 34个碱基对 - tRNA1Gln - 15个碱基对 - tRNAmMet - 47个碱基对 - tRNA2Gln - 37个碱基对 - tRNA2Gln。编码tRNAmMet、tRNA1Gln和tRNA2Gln的重复基因具有相同的序列,这与先前用tRNA分子确定的序列相同。这些tRNA序列之前有一个单一的启动子区域,在转录起始位点上游七个碱基对处存在一个“Pribnow框”序列。分隔七个tRNA序列的间隔区在大小和核苷酸序列上彼此不同。讨论了这些序列对前体加工的可能影响。最初在λpsu°2 DNA中鉴定并显示包含七个tRNA基因的一个限制片段已在大肠杆菌染色体中检测到,从而表明该tRNA基因簇以与转导噬菌体基因组相同的组织形式存在于细菌基因组中。