エレクトロポレーションによる Agrobacterium rhizogenes MAFF03-01724株の形質転換

概要

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Agrobacterium rhizogenes MAFF03-01724 (mikimopine type), isolated in Japan, showed resistance to ampicillin, carbenicillin, chloramphenicol, kanamycin, rifampicin, streptomycin, tetracycline and vancomycin. Two kanamycin sensitive mutants (Km^s) and two chloramphenicol sensitive ones (Cm^s) were obtained from strain 1724 by treatment with N-metyl-N'-nitro-N-nitrosoguanidine (NTG) for transformation by transfer of foreign genes possessing antibiotic selection markers. Transformants were obtained from the Km^s mutant by electroporation with binary vector, pBIN19, and/or pF 11 constructed by cloning of BamHI fragment (8.5 kb) of pRi1724 into pHSG298 vector. To improve transformation frequency, strain 1724 was cultured in a flask with bames and cells washed with 10% glycerol by repeated pipetting in extrusion. By this modified preparation, transformation frequency of pBIN19 increased fifty times.
日本植物病理学会の論文
1993-10-25