Dobrovol'ski P, Sakanian V A
Genetika. 1986 Nov;22(11):2693-701.
Hydroxylamine-induced mutants of the plasmid pPD6 (8.4 kb) were isolated which are resistant to high doses of tetracycline. One of the plasmids studied--pPD21 is a multicopy mutant, another one, pPD12 is a dimeric form of the pPD6 plasmid. The pPD12 plasmid is very unstable, its derivative, pPD13 spontaneous mutant acquiring stability but not the ability to resolve DNA multimeric forms into monomeric forms. Multicopy bireplicon pPD619 plasmid was constructed by joining in vitro pPD6 and pUC19 plasmids. Removing the replicon pUC19 from the bireplicon plasmid gives a new low-copy plasmid pPD620. All of the plasmids constructed were mobilized by the conjugative pRK2013 plasmid into the strains of Escherichia coli, Pseudomonas aeruginosa and Agrobacterium tumefaciens. The pPD6 plasmid and its derivatives can be used as cloning vectors.
分离出了羟胺诱导的质粒pPD6（8.4 kb）突变体，这些突变体对高剂量四环素具有抗性。所研究的其中一种质粒——pPD21是多拷贝突变体，另一种质粒pPD12是pPD6质粒的二聚体形式。pPD12质粒非常不稳定，其衍生物pPD13自发突变体获得了稳定性，但没有将DNA多聚体形式解析为单体形式的能力。通过体外连接pPD6和pUC19质粒构建了多拷贝双复制子pPD619质粒。从双复制子质粒中去除复制子pUC19可得到一种新的低拷贝质粒pPD620。构建的所有质粒都通过接合型pRK2013质粒转移到大肠杆菌、铜绿假单胞菌和根癌土壤杆菌菌株中。pPD6质粒及其衍生物可用作克隆载体。
