Phillips-Jones M K
Department of Molecular Biology and Biotechnology, University of Sheffield, UK.
FEMS Microbiol Lett. 1993 Feb 1;106(3):265-70. doi: 10.1111/j.1574-6968.1993.tb05974.x.
To determine whether bacterial luciferase is expressed in the anaerobe Clostridium perfringens to produce an oxygen-requiring bioluminescence reaction, a suitable plasmid vector possessing the luxA and luxB genes of Vibrio fischeri was constructed and introduced into C. perfringens cells. luxAB were placed under the transcriptional control of the C. perfringens alpha-toxin gene promoter region. Suitable ribosome binding sites were introduced upstream of both genes. Bioluminescence was strongly expressed in C. perfringens transformants. Comparisons of in vivo and in vitro bioluminescence measurements demonstrated that in vivo data constituted a quantitative measure of gene expression. This is the first study to show that luxA and luxB genes can be expressed in an anaerobic bacterium and that bioluminescence can be used as a quantitative reporter system in future in vivo studies of gene expression in C. perfringens.
为了确定细菌荧光素酶是否在厌氧菌产气荚膜梭菌中表达以产生需氧生物发光反应，构建了一个携带费氏弧菌luxA和luxB基因的合适质粒载体，并将其导入产气荚膜梭菌细胞。luxAB置于产气荚膜梭菌α-毒素基因启动子区域的转录控制之下。在两个基因的上游引入了合适的核糖体结合位点。在产气荚膜梭菌转化体中强烈表达生物发光。体内和体外生物发光测量的比较表明，体内数据构成了基因表达的定量指标。这是第一项表明luxA和luxB基因可在厌氧菌中表达且生物发光可在未来产气荚膜梭菌基因表达的体内研究中用作定量报告系统的研究。
