construction of recombinant vectors containing clavulanic acid antibiotic regulatory gene, clar, isolated of streptomyces clavuligerus strains
نویسندگان
چکیده
the clar of streptomyces clavuligerus in the clavulanic acid gene cluster encodes a transcriptional regulator that controls clavulanic acid biosynthesis. the main goal of this study was isolation and molecular detection of the clar gene and its cloning in the streptomyces specific vector (pma:: hyg). by cinsideration of the clar gene’s start codon, the specific primers were designed. after genomic dna extraction from s. clavuligerus, the clar gene was amplified by polymerase chain reaction (pcr). the structure of the amplified clar was confirmed by nested-pcr, pcr-restriction fragment length polymorphism (pcr-rflp), and sequencing. a ligation mixture was prepared with the isolated clar gene and cut pma::hyg vector. escherichia coli competent cells were finally transformed with the ligation mixture. presence of the recombinant vector in the transformed colonies was then confirmed by the colony-pcr procedure. the clar gene was also isolated from s. clavuligerus dsm41826, cloned and sequenced in the same manner. the pma::hyg vector is a shuttle vector, which exists as a multicopy plasmid in e. coli, and as an integrative plasmid in streptomyces. therefore, the newly constructed vectors of this study can be regarded as an appropriate tool for site-directed mutagenesis and gene replacement strategies in s. clavuligerus.
منابع مشابه
Construction of Recombinant Vectors Containing Clavulanic Acid Antibiotic Regulatory Gene, claR, Isolated of Streptomyces clavuligerus Strains
The claR of Streptomyces clavuligerus in the clavulanic acid gene cluster encodes a transcriptional regulator that controls clavulanic acid biosynthesis. The main goal of this study was isolation and molecular detection of the claR gene and its cloning in the Streptomyces specific vector (pMA:: hyg). By cinsideration of the claR gene’s start codon, the specific primers were designed. After geno...
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متن کاملmolecular analysis of the clavulanic acid regulatory gene isolated from an iranian strain of streptomyces clavuligerus, ptcc 1709
objective: the clavulanic acid regulatory gene (clar) is in the clavulanic acid biosynthetic gene cluster that encodes clar. this protein is a putative regulator of the late steps of clavulanic acid biosynthesis. the aim of this research is the molecular cloning of clar, isolated from the iranian strain of streptomyces clavuligerus (s. clavuligerus). materials and methods: in this experimental ...
متن کاملMolecular Analysis of the Clavulanic Acid Regulatory Gene Isolated from an Iranian Strain of Streptomyces Clavuligerus , PTCC 1709
OBJECTIVE The clavulanic acid regulatory gene (claR) is in the clavulanic acid biosynthetic gene cluster that encodes ClaR. This protein is a putative regulator of the late steps of clavulanic acid biosynthesis. The aim of this research is the molecular cloning of claR, isolated from the Iranian strain of Streptomyces clavuligerus (S. clavuligerus). MATERIALS AND METHODS In this experimental ...
متن کاملOverproduction of Clavulanic Acid by UV Mutagenesis of Streptomyces clavuligerus
Clavulanic acid is produced industrially by fermentation of Streptomyces clavuligerus and researches have increased its production by strain improvement, recombinant DNA technology, and media composition and growth condition optimization. The main objective of this study was to increase the level of clavulanic acid production from Streptomyces clavuligerus (DSM 738), using UV irradiation. After...
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عنوان ژورنال:
iranian journal of biotechnologyناشر: national institute of genetic engineering and biotechnology
ISSN 1728-3043
دوره 7
شماره 2 2009
کلمات کلیدی
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