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[R6K质粒DNA介导的肠道细菌转化]

[Entobacterial transformation by R6K plasmid DNA].

作者信息

Gnedoĭ S N, Babushkina L M, Frolov V N, Levashev V S

出版信息

Zh Mikrobiol Epidemiol Immunobiol. 1977(1):99-104.

PMID:320804
Abstract

Preparations of DNA of R6K plasmide obtained by various methods on the basis of the "clarified" lysate: by gel filtration on sepharose 4B, centrifugation in cesium chloride with ethidium bromide gradient, were analysed in the E. coli C600. S. typhimurium AG37, Pr. vulgaris 4636, S. marcescens 20-10 transformation. The frequency of transformation proved to depend on the extent of DNA purification. Factors influencing the E. coli C600 competence and the transformation efficacy (the phase of the culture growth, the concentration of cells in the mixture with DNA, theCaCl2 concentration, the time of the cell incubation at 42 degrees C) were studied. Kinetics of the phenotypical expression of the plasmide signs of ampicillin and streptomycin resistance in transformation was also studied in this work.

摘要

基于“澄清”裂解物,通过多种方法获得的R6K质粒DNA制剂:通过在琼脂糖4B上进行凝胶过滤、在氯化铯-溴化乙锭梯度中离心,在大肠杆菌C600、鼠伤寒沙门氏菌AG37、普通变形杆菌4636、粘质沙雷氏菌20-10转化中进行分析。转化频率被证明取决于DNA纯化程度。研究了影响大肠杆菌C600感受态和转化效率的因素(培养物生长阶段、与DNA混合时细胞的浓度、氯化钙浓度、细胞在42摄氏度下的孵育时间)。在这项工作中还研究了转化中氨苄青霉素和链霉素抗性的质粒标志的表型表达动力学。

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