A pair of mobilizable shuttle vectors conferring resistance to spectinomycin for molecular cloning in Escherichia coli and in gram-positive bacteria

We previously designed a vector strategy that allows transfer by conjugation of recombinant plasmids from Escherichia coli to various Gram-positive bacilli and cocci. We report here the construction of plasmid vectors which exploit this broad host range transfer system. These vectors, pAT28 and pAT29, are composed of 1) the origins of replication of pUC and of the broad host range enterococcal plasmid pAM beta 1; 2) a spectinomycin resistance gene expressed in gram-negative and in gram-positive bacteria; 3) the origin of transfer of the IncP plasmid RK2; and 4) the multiple cloning site and the lacZ alpha reporter gene of pUC18 (pAT28) and pUC19 (pAT29). These plasmids can be efficiently mobilized by any self-transferable IncP plasmid co-resident in E. coli donors.