, Volume 40, Issue 3, pp 257-262

Expression ofEscherichia coli recA andada genes inSaccharomyces cerevisiœ using a vector with geneticin resistance

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Abstract

Construction ofE. coli-yeast shuttle plasmids containing theneo selection gene is described. The protein-coding regions of theE. coli ada orrecA genes under the control of theADH1 promoter and terminator were ligated into theSphI unique site of pNF2 to produce pMSada and pMSrecA, respectively. The plasmids were used for transformation of the haploid and diploidpso4-1 strains ofS. cerevisiœ and their corresponding wild types. Transformants were obtained by selection for geneticin (G418) resistance. Crude protein samples were extracted from the individual transformants. Both the RecA and Ada proteins were present in all strains containing therecA andada genes on plasmids, respectively. Thus the geneticin selection system was successfully used for the preparation of model, yeast strains.