Chakrabarty A M, Mylroie J R, Friello D A, Vacca J G
Proc Natl Acad Sci U S A. 1975 Sep;72(9):3647-51. doi: 10.1073/pnas.72.9.3647.
Conditions optimal for the transformation of Pseudomonas putida and E. coli with a drug-resistance factor (RP 1) DNA, which specifies resistance to carbenicillin, tetracycline, kanamycin, and neomycin, are described. The transformants retain all the fertility, incompatibility, and drug-resistance characteristics present in the parent. Covalently-closed circular molecules of almost identical contour lengths have been isolated from the parent and the transformants. The frequency of transformation is drastically reduced by treatment of RP 1 DNA with DNase and by denaturation or sonication. Shearing of RP 1 DNA in vitro and their subsequent introduction in P. putida cells, by transformation, produces transformants that exhibit a wide range of drug-resistant phenotypes, including those which are resistant to neomycin but sensitive to kanamycin. Isolation of such neomycin-resistant but kanamycin-sensitive transformants indicates that there might be two separate mechanisms specified by RP 1 for resistance to the two antibiotics.
本文描述了用一种对羧苄青霉素、四环素、卡那霉素和新霉素具有抗性的抗药因子（RP 1）DNA转化恶臭假单胞菌和大肠杆菌的最佳条件。转化体保留了亲代中存在的所有育性、不相容性和抗药特性。已从亲代和转化体中分离出轮廓长度几乎相同的共价闭合环状分子。用DNase处理RP 1 DNA以及变性或超声处理会大幅降低转化频率。在体外剪切RP 1 DNA并随后通过转化将其导入恶臭假单胞菌细胞中，会产生表现出广泛抗药表型的转化体，包括对新霉素有抗性但对卡那霉素敏感的转化体。分离出这种对新霉素有抗性但对卡那霉素敏感的转化体表明，RP 1可能存在两种独立的机制来对抗这两种抗生素。
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