OPTIMIZATION OF STREPTOMYCES-AUREOFACIENS TRANSFORMATION AND DISRUPTION OF THE HRDA GENE ENCODING A HOMOLOG OF THE PRINCIPAL SIGMA-FACTOR

被引:24
作者
KORMANEC, J
REZUCHOVA, B
FARKASOVSKY, M
机构
[1] Institute of Molecular Biology, Slovak Academy of Sciences, 842 51 Bratislava
来源
JOURNAL OF GENERAL MICROBIOLOGY | 1993年 / 139卷
关键词
D O I
10.1099/00221287-139-10-2525
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Conditions for the preparation of protoplasts from Streptomyces aureofaciens and transformation with several replicative and integrative plasmids were optimized. Of all the replicative plasmids used, carrying various Streptomyces replication origins, only plasmid pGM9 produced transformants, with a transformation efficiency of 2 x 10(3) (mug DNA)-1. When plasmid DNA isolated from S. aureofaciens was used, the transformation efficiency increased to 10(5) transformants (pg DNA)-1. Transformation of protoplasts with integrative plasmids containing part of the hrdA gene gave rise to transformants with an efficiency of approximately 30 transformants (mug DNA)-1. Integration frequency was strongly reduced when plasmid DNA was modified by dam or dcm methylase. By integrative transformation, via a double crossover, a stable null mutant of the hrdA gene was prepared. This mutation appeared to have no obvious effect on growth, morphology, differentiation and production of chlortetracycline.
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收藏
页码:2525 / 2529
页数:5
相关论文
共 24 条
[1]  
BALTZ RH, 1981, J GEN MICROBIOL, V127, P137
[2]   RNA-POLYMERASE HETEROGENEITY IN STREPTOMYCES-COELICOLOR A3(2) [J].
BUTTNER, MJ .
MOLECULAR MICROBIOLOGY, 1989, 3 (11) :1653-1659
[3]   CONSTRUCTION AND CHARACTERIZATION OF STREPTOMYCES-COELICOLOR A3(2) MUTANTS THAT ARE MULTIPLY DEFICIENT IN THE NONESSENTIAL HRD-ENCODED RNA-POLYMERASE SIGMA FACTORS [J].
BUTTNER, MJ ;
LEWIS, CG .
JOURNAL OF BACTERIOLOGY, 1992, 174 (15) :5165-5167
[4]   MUTATIONAL CLONING IN STREPTOMYCES AND THE ISOLATION OF ANTIBIOTIC PRODUCTION GENES [J].
CHATER, KF ;
BRUTON, CJ .
GENE, 1983, 26 (01) :67-78
[5]  
CHATER KF, 1986, BACTERIA, V9, P119
[6]   THE STREPTOMYCES-COELICOLOR-WHIB GENE ENCODES A SMALL TRANSCRIPTION FACTOR-LIKE PROTEIN DISPENSABLE FOR GROWTH BUT ESSENTIAL FOR SPORULATION [J].
DAVIS, NK ;
CHATER, KF .
MOLECULAR & GENERAL GENETICS, 1992, 232 (03) :351-358
[7]   A VERSATILE SHUTTLE COSMID VECTOR FOR USE IN ESCHERICHIA-COLI AND ACTINOMYCETES [J].
DENIS, F ;
BRZEZINSKI, R .
GENE, 1992, 111 (01) :115-118
[8]   STRUCTURE AND FUNCTION OF BACTERIAL SIGMA FACTORS [J].
HELMANN, JD ;
CHAMBERLIN, MJ .
ANNUAL REVIEW OF BIOCHEMISTRY, 1988, 57 :839-872
[9]   GENE DISRUPTION AND GENE REPLACEMENT IN STREPTOMYCES VIA SINGLE-STRANDED-DNA TRANSFORMATION OF INTEGRATION VECTORS [J].
HILLEMANN, D ;
PUHLER, A ;
WOHLLEBEN, W .
NUCLEIC ACIDS RESEARCH, 1991, 19 (04) :727-731
[10]  
Hopwood D. A., 1985, GENETIC MANIPULATION