THE CONSTRUCTION OF A REPORTER SYSTEM AND USE FOR THE INVESTIGATION OF CLOSTRIDIUM-PERFRINGENS GENE-EXPRESSION

被引:11
作者
BULLIFENT, HL
MOIR, A
TITBALL, RW
机构
[1] CHEM & BIOL DEF ESTAB,SALISBURY SP4 0JQ,WILTS,ENGLAND
[2] UNIV SHEFFIELD,DEPT MOLEC BIOL & BIOTECHNOL,SHEFFIELD S10 2TN,S YORKSHIRE,ENGLAND
关键词
CLOSTRIDIUM PERFRINGENS; RECOMBINANT DNA; CHLORAMPHENICOL ACETYLTRANSFERASE; SHUTTLE VECTOR; TOXIN;
D O I
10.1111/j.1574-6968.1995.tb07761.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A reporter system was constructed to enable the study of gene expression in Clostridium perfringens. The system was based on plasmid shuttle vector pJIR410, which contained the C. perfringens erythromycin resistance gene. The vector was modified by the introduction of a DNA fragment comprising the open reading frame of the C. perfringens chloramphenicol acetyltransferase gene and flanking transcriptional terminators. The presence of a unique restriction site, engineered into the extreme 5' end of the open reading frame enabled a promoter region to be inserted to form an in-frame transcriptional fusion with catP. The system was tested by inserting the promoter region of the alpha-toxin gene of C. perfringens. The production of chloramphenicol acetyltransferase in C. perfringens was monitored during growth and the pattern of expression was shown to reflect levels of pie mRNA and alpha-toxin in the parent strain.
引用
收藏
页码:99 / 105
页数:7
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