枯草芽孢杆菌色氨酰 - tRNA合成酶结构基因的克隆及核苷酸序列

Cloning and nucleotide sequence of the structural gene coding for Bacillus subtilis tryptophanyl-tRNA synthetase.

作者信息

Chow K C, Wong J T

机构信息

Department of Biochemistry, University of Toronto, Ont., Canada.

出版信息

Gene. 1988 Dec 20;73(2):537-43. doi: 10.1016/0378-1119(88)90518-5.

DOI:10.1016/0378-1119(88)90518-5

PMID:3149612

Abstract

A 1.47-kb DNA fragment that carries the tryptophanyl-tRNA synthetase (TrpRS) gene of Bacillus subtilis has been cloned into the pUC8 plasmid. The recombinant plasmid, pTSQ2, conferred temperature-resistance to the temperature-sensitive trpS ts mutant of B. subtilis through chromosomal transformation, and to that of Escherichia coli through complementation. The pTSQ2 could be stably maintained in E. coli DH5 alpha, causing in the host cell a 200-fold amplification of TrpRS activity. The complete nucleotide sequence of the cloned fragment has been determined. A putative transcriptional promoter, a Shine-Dalgarno sequence, the 990-bp trpS gene proper, as well as a transcriptional terminator have been identified.

摘要

一段携带枯草芽孢杆菌色氨酰 - tRNA合成酶（TrpRS）基因的1.47 kb DNA片段已被克隆到pUC8质粒中。重组质粒pTSQ2通过染色体转化赋予枯草芽孢杆菌温度敏感型trpS ts突变体温度抗性，并通过互补作用赋予大肠杆菌温度抗性。pTSQ2能够在大肠杆菌DH5α中稳定维持，使宿主细胞中的TrpRS活性放大200倍。已确定克隆片段的完整核苷酸序列。已鉴定出一个推定的转录启动子、一个Shine - Dalgarno序列、990 bp的trpS基因本身以及一个转录终止子。