Open AccessHigh efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA‐restricting streptomycetes
Author(s) -
Flett Fiona,
Mersinias Vassilios,
Smith Colin P.
Publication year - 1997
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1997.tb13882.x
Subject(s) - streptomyces coelicolor , plasmid , biology , escherichia coli , genetics , dna , methylation , dna methylation , bacteria , streptomyces , gene , microbiology and biotechnology , gene expression
Many streptomycetes, including S. coelicolor A3(2), possess a potent methyl‐specific restriction system which can present an effective barrier to the introduction of heterologous DNA. We have compared the efficiency of intergeneric conjugal transfer of different types of plasmids to S. coelicolor and S. lividans 66 using two E. coli donors: the standard, methylation proficient strain S17‐1. and the methylation deficient donor, ET12567(pUB307). We demonstrate that the methylation deficient donor can yield > 10 4 ‐fold more S. coelicolor exconjugants than the standard donor. In the case of pSET152 derivatives, which integrate into the host chromosome by site‐specific recombination, up to 10% of streptomycete spores in the conjugation mixture inherit the plasmid. The conjugation procedure is efficient enough to obtain exconjugants with ‘suicide’ delivery plasmids and therefore provides a simple route for conducting gene disruptions in methyl DNA‐restricting streptomycetes, and possibly other bacteria.