CLONING OF THE WIDE SPECTRUM AMIDASE GENE FROM BREVIBACTERIUM SP R312 BY GENETIC COMPLEMENTATION - OVEREXPRESSION IN BREVIBACTERIUM SP AND ESCHERICHIA-COLI

被引:4
作者
AZZA, S [1 ]
BIGEY, F [1 ]
ARNAUD, A [1 ]
GALZY, P [1 ]
机构
[1] ECOLE NATL SUPER AGRON MONTIPELLIER, CHAIR MICROBIOL IND & GENET MICOORGANISMES, F-34060 MONTPELLIER 1, FRANCE
关键词
BREVIBACTERIUM SP; AMIE GENE CLONING; COMPLEMENTATION AND EXPRESSION;
D O I
10.1111/j.1574-6968.1994.tb07155.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The amiE gene of Brevibacterium sp. R312 encoding wide spectrum amidase was isolated by complementation of a Brevibacterium sp. mutant using a plasmid gene bank of chromosomal DNA. The amiE structural gene and its promoter were localized on a 1.8-kb fragment by subsequent subcloning and complementation studies. Another promoter localized in the pSR 1 fragment of the cloning vector was shown to be able to control amiE gene expression. In Brevibacterium sp., the investigation of amidase activities related to one copy of the gene suggested that the regulation of the amiE gene expression was under negative control. High expression levels have been obtained in Brevibacterium sp. and, after substitution of the amiE promoter by the tac promoter, in Escherichia coli.
引用
收藏
页码:129 / 136
页数:8
相关论文
共 23 条
[21]   DETECTION OF SPECIFIC SEQUENCES AMONG DNA FRAGMENTS SEPARATED BY GEL-ELECTROPHORESIS [J].
SOUTHERN, EM .
JOURNAL OF MOLECULAR BIOLOGY, 1975, 98 (03) :503-+
[22]   THE DETERMINATION OF SMALL QUANTITIES OF BACTERIA BY MEANS OF THE BIURET REACTION [J].
STICKLAND, LH .
JOURNAL OF GENERAL MICROBIOLOGY, 1951, 5 (04) :698-703
[23]   PURIFICATION AND PROPERTIES OF AN ACYLAMIDE AMIDOHYDROLASE (E.C.3.5.1.4) WITH A WIDE ACTIVITY SPECTRUM FROM BREVIBACTERIUM-SP R 312 [J].
THIERY, A ;
MAESTRACCI, M ;
ARNAUD, A ;
GALZY, P ;
NICOLAS, M .
JOURNAL OF BASIC MICROBIOLOGY, 1986, 26 (05) :299-311