RESUMEN
Mobilizable shuttle plasmids containing the origin of transfer (oriT) region of plasmid F (IncFI), ColIb-P9 (IncI1), and RP4/RP1 (IncPalpha) were constructed to test the ability of the cognate conjugation system to mediate gene transfer from Escherichia coli to Streptomyces. The conjugative system of the IncPalpha plasmids was shown to be most effective in conjugative transfer, giving peak values of (2.7 +/- 0.2) x 10(-2) S. lividans TK24 exconjugants per recipient cell. To assess whether the mating-pair formation system or the DNA-processing apparatus of the IncPalpha plasmids is crucial in conjugative transfer, an assay with an IncQ-based mobilizable plasmid (RSF1010) specifying its own DNA-processing system was developed. Only the IncPalpha plasmid mobilized the construct to S. lividans indicating that the mating-pair formation system is primarly responsible for the promiscuous transfer of the plasmids between E. coli and Streptomyces. Dynamic of conjugative transfer from E. coli to S. lividans was investigated and exconjugants starting from the first hour of mating were obtained.
Asunto(s)
Conjugación Genética , Escherichia coli/genética , Streptomyces/genética , Antibacterianos/biosíntesis , Factor F/genética , Streptomyces/metabolismo , Streptomyces lividans/genética , Streptomyces lividans/metabolismoRESUMEN
Streptomyces globisporus strains with knockouts in lndF and lndL genes, previously identified as possibly encoding cyclases governing cyclization of the nascent oligoketide ('polyketide') chain during the biosynthesis of the antitumor angucycline landomycin E, were prepared. On combining the results of sequence analysis and HPLC of extracts from mutant strains, lndL was suggested to control the first cyclization-aromatization event and lndF to be responsible for the 3rd-4th ring formation.
Asunto(s)
Aminoglicósidos/biosíntesis , Enzimas/genética , Genes Bacterianos/fisiología , Mutagénesis Insercional , Streptomyces/enzimología , Streptomyces/genética , Aminoglicósidos/química , Proteínas Bacterianas/genética , Southern Blotting , Cromatografía Líquida de Alta Presión , Cromatografía en Capa Delgada , Enzimas/metabolismo , Streptomyces/químicaRESUMEN
The regulatory genes lanI and lndI have been cloned from the landomycin A (LaA) producer Streptomyces cyanogenus S136 and from the landomycin E (LaE) producer Streptomyces globisporus 1912, respectively and both have been sequenced. A culture of S. globisporus I2-1 carrying a disrupted lndI gene did not produce LaE and other related intermediates. Complementation of S. globisporus I2-1 with either the lndI or lanI gene reconstituted LaE production indicating that LanI and LndI are involved in activation of structural genes in the respective clusters. Structural features of these regulatory genes are discussed.