Development and use of a conditional antisense mutagenesis system in mycobacteria

被引:57
作者
Parish, T [1 ]
Stoker, NG [1 ]
机构
[1] UNIV LONDON LONDON SCH HYG & TROP MED,DEPT CLIN SCI,LONDON WC1E 7HT,ENGLAND
基金
英国惠康基金;
关键词
vector; acetamidase; histidine; Mycobacterium; antisense;
D O I
10.1016/S0378-1097(97)00317-0
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Expression from a 2.3 kb region upstream of the inducible acetamidase gene from Mycobacterium smegmatis was shown to be upregulated by acetamide. A DNA fragment containing the start of the M. smegmatis hisD gene was cloned in front of the promoter, such that the antisense message was produced. When this construct was induced in vivo, the bacteria became phenotypically histidine auxotrophs; this auxotrophy was restored by histidine supplementation. Auxotrophy was not observed under non-induced conditions. Antisense mutagenesis may be useful for observing the phenotypic inactivation of specific mycobacterial genes, and an inducible system such as that described would allow the study of essential genes.
引用
收藏
页码:151 / 157
页数:7
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