The <i>Streptomyces aureofaciens</i> plasmid pIMB R8 and its use for shuttle vector construction

Godány, Andrej; Lacová, Bibiána; Zelinka, J.

doi:10.1002/jobm.3620301007

Cited by 6 publications

(1 citation statement)

References 19 publications

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“…There were two reasons for using ethidium bromide as an agent for plasmid curing from S. aureofaciens transformants. The first one was the high stability of the plasmid replication in S. aureofaciens ( GODANY et al 1990) and the second one was the good efficiency of ethidium bromide as mutagen.…”

Section: Discussionmentioning

confidence: 99%

Highly transformable mutants of Streptomyces aureofaciens containing restriction‐modification systems

Muchová

Lacová

Godány

et al. 1991

J. Basic Microbiol.

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Streptomyces aureofaciens 13 is a mutant defective in chlortetracycline production. It was chosen as a potentially useful host for gene cloning in investigations of the organization of the biosynthetic genes for the tetracycline antibiotic pathway. From the Streptomyces aureofaciens 13 strain, three suitable clones were used for our work. The conditions for optimal formation and efficient transformation of protoplasts with plasmid DNAs have been determined. Transformation frequencies of about 10(4) to 10(5) per microgram of plasmid DNA were obtained when plasmids were isolated from Streptomyces strains. From the patterns of restriction enzyme digestion of plasmid DNA isolated from Streptomyces aureofaciens transformants, it was observed that the clones express modification systems which render plasmid DNAs resistant to cleavage by HindIII and EcoRI. Additionally, one of the clones produces the restriction endonuclease Sau13I (isoschizomer of SauI). The presence of the restriction-modification system of Sau13I does not reduce the efficiency of plasmid transformation.

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Section: Discussionmentioning

confidence: 99%

Highly transformable mutants of Streptomyces aureofaciens containing restriction‐modification systems

Muchová

Lacová

Godány

et al. 1991

J. Basic Microbiol.

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Streptomyces aureofaciens strains as hosts for cloning of genes affecting antibiotic production

et al. 1991

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Analysis of the Site-Specific Integration System of the Streptomyces aureofaciens Phage μ1/6

Farkašovská

Godány

2011

Curr Microbiol

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The bacteriophage μ1/6 integrates its DNA into the chromosome of tetracycline producing strains of Streptomyces aureofaciens by a site-specific recombination process. A bioinformatic analysis of the μ1/6 genome revealed that orf5 encodes a putative integrase, a basic protein of 416 amino acids. The μ1/6 integrase was found to belong to the integrase family of site-specific tyrosine recombinases. The phage attachment site (attP) was localized downstream of the int gene. The attachment junctions (attL and attR) were determined, allowing identification of the bacterial attachment site (attB). All attachment sites shared a 46-bp common core sequence within which a site-specific recombination occurs. This core sequence comprises the 3' end of a putative tRNA(Thr) gene (anticodon TGT) which is completely restored in attL after integration of the phage into the host genome. An integration vector containing μ1/6 int-attP region was inserted stably into the S. aureofaciens B96, S. lividans TK24, and S. coelicolor A3. The μ1/6 integrase was shown to be functional in vivo in heterologous Escherichia coli without any other factors encoded by Streptomyces. In vitro recombination assay using purified μ1/6 integrase demonstrated its ability to catalyze integrative recombination in the presence of a crude extract of E. coli cells.

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The Streptomyces aureofaciens plasmid pIMB R8 and its use for shuttle vector construction

Cited by 6 publications

References 19 publications

Highly transformable mutants of Streptomyces aureofaciens containing restriction‐modification systems

Highly transformable mutants of Streptomyces aureofaciens containing restriction‐modification systems

Streptomyces aureofaciens strains as hosts for cloning of genes affecting antibiotic production

Analysis of the Site-Specific Integration System of the Streptomyces aureofaciens Phage μ1/6

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