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Efficient transformation of Mesorhizobium huakuii subsp. rengei and Rhizobium species
Authors:Hayashi M  Maeda Y  Hashimoto Y  Murooka Y
Affiliation:Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka 565-0871, Japan.
Abstract:The transformation of Mesorhizobium huakuii subsp. rengei B3 with either pBBR122 or pKT230 was carried out. We determined the optimal conditions required for transformation by electroporation and obtained up to 10(5) CFU/microg pBBR122. Plasmids prepared from strain B3 yielded higher transformation efficiency than those from various dam or dcm mutant strains of Escherichia coli. This result suggests that a high transformation efficiency is not related to the methylation of plasmid DNA in E. coli. Using the optimal conditions for electroporation, we performed transformation of several species of Rhizobium, Mesorhizobium and Sinorhizobium. All tested strains of these species were transformed with pBBR122. Strains of M. huakuii and R. phaseoli AHU1133 were transformed at high efficiency, whereas transformation efficiencies of Rhizobium sp. NGR234 and S. meliloti strains were less than 2 x 10(3) CFU/microg plasmid DNA.
Keywords:transformation  Mesorhizobium huakuii subsp  rengei B3  Astragalus sinicus  Rhizobium species
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