Open AccessPlasmid transfer by conjugation from Escherichia coli to Gram‐positive bacteria
Author(s) -
TrieuCuot P.,
Carlier C.,
Martin P.,
Courvalin P.
Publication year - 1987
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.1574-6968.1987.tb02558.x
Subject(s) - plasmid , escherichia coli , microbiology and biotechnology , biology , enterococcus faecalis , kanamycin , transformation (genetics) , shuttle vector , bacteria , listeria monocytogenes , plasmid preparation , streptococcus agalactiae , pbr322 , streptococcus , dna , recombinant dna , genetics , gene , vector (molecular biology) , antibiotics
Abstract We have developed a vector strategy that allows transfer of plasmid DNA by conjugation from Escherichia coli to various Gram‐positive bacteria in which transformation via natural competence has not been demonstrated. The prototype vector constructed, pAT187, contains the origins of replication of pBR322 and of the broad host range streptococcal plasmid pAMβ1, a kanamycin resistance gene known to be expressed in both Gram‐negative and Gram‐positive bacteria, and the origin of transfer of the IncP plasmid RK2. This shuttle plasmid can be mobilised efficiently by the self‐transferable IncP plasmid pRK212.1 co‐resident in the E. coli donors, and was successfully transferred by filter matings at frequencies of 2 × 10 −8 to 5 × 10 −7 to Enterococcus faecalis, Streptococcus lactis, Streptococcus agalactiae, Bacillus thuringiensis, Listeria monocytogenes and Staphylococcus aureus .