[Actinomycete plasmid and integrative vectors based on DNA of the temperate Phi C31 actinophage, determining limitation of lytic development of phage Phi C31, not dependent on repressor].
Bireplicon plasmids were constructed. The plasmids consist of DNAs of the streptomycete plasmid pIJ702, the Escherichia coli plasmid pUC19 and the phi C31 actinophage genome fragment encoding the function of the site-specific integration into the chromosome. Part of the plasmids transformed Streptomyces lividans TK64 to thiostreptone resistance. The DNA transforming activity depended on the mutual orientations of the blocks used for the construction and probably depended on the structural stability of the plasmids in S. lividans. The integrative vectors consisting of the pUC19 plasmid DNA and the phage genome fragment with the integrative function efficiently transformed S. lividans. No phage plagues were detected with the standard procedure of integrants' infection by phi C31 phage, despite the absence of the phi C31 phage repressor gene in the integrated DNA. The phi C31 phage mutants including clear and virulent ones were not capable of lytic growth on the integrants. The region determining the limitation of the phi C31 phage lytic development was localized by the deletion analysis of the bireplicon plasmids. As a result actinomycete monoreplicon plasmids were formed. The region is the 1.3 kb phage fragment whose right end maps at 0.2 kb preceding the right end of the phi C31 phage genome linear map.
Sladkova
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IA
Orekhov
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AV
D001433:Bacteriolysis / Q000502:physiology*; D001435:Bacteriophages / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics / Q000821:virology; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics*; D012097:Repressor Proteins / Q000235:genetics*; D013302:Streptomyces / Q000235:genetics* / Q000821:virology; D014170:Transformation, Genetic
D004279:DNA, Viral; D012097:Repressor Proteins
Aktinomitsetnye plazmidnye i integrativnye vektory na osnove DNK umerennogo aktinofaga Phi C31, opredeliaiushchie organichenie liticheskogo razvitiia faga Phi C 31, ne zavisimoe ot repressora.