Construction of Efficient Conjugal Plasmids Between Escherichia coli and Streptomycetes
大肠杆菌-链霉菌高效接合载体的构建及其应用

Conjugal plasmid pGH112 has been developed based on the replicons of Streptomyces coelicolor plasmid SCP2 * and E.coli ColE. The plasmid contains ampicilin resistance gene( amp ) for selection in E.coli and thiostrepton resistance gene ( tsr ) for selection in Streptomycetes, and a 0.76 kb oriT fragment of (IncP) RK2. Conjugal transfer of pGH112 was performed from E.coli to S.coelicolor A3(2), S.avermitilis , S.lividans TK54, S.toxytricini NNRL15443, S.venezuelae ISP5230 and Sacc.erythraea by conjugation, results show that the plasmid was able to transfer efficenctly from E.coli to Streptomycetes, was stably inherited in the recipients. pGH113 was constructed from pGH112 by combining the constitutive erm E * promoter with green fluorescent protein gene( gfp ).