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枯草芽孢杆菌中质粒转化的特性:动力学性质及DNA构象的影响

Characterization of plasmid transformation in Bacillus subtilis: kinetic properties and the effect of DNA conformation.

作者信息

Contente S, Dubnau D

出版信息

Mol Gen Genet. 1979 Jan 2;167(3):251-8. doi: 10.1007/BF00267416.

Abstract

Transformation of competent cells of Bacillus subtilis with antibiotic resistance plasmid DNA has shown that (a) competence for plasmid and chromosomal DNA develops with similar kinetics; (b) DNA linearized with a variety of restriction endonucleases does not transform; (c) CCC plasmid DNA is inactivated for transformation by a single nick; (d) T4 ligase restores transforming activity to both nicked and linearized DNA; (E) CCC relaxed DNA is fully active in transformation; (f) the DNA concentration-dependence of plasmid transformation is first order; and (g) plasmid transformation proceeds with a low efficiency, requiring the uptake of 10(3) to 10(4) DNA molecules per transformant. Based on this information, a model for the processing of chromosomal, plasmid and transfecting DNA is proposed.

摘要

用抗生素抗性质粒DNA对枯草芽孢杆菌感受态细胞进行转化的实验表明:(a)质粒DNA和染色体DNA的感受态形成动力学相似;(b)用多种限制性内切酶线性化的DNA不能转化;(c)单一切口会使CCC质粒DNA失去转化活性;(d)T4连接酶可恢复切口DNA和线性化DNA的转化活性;(e)CCC松弛型DNA在转化中具有完全活性;(f)质粒转化的DNA浓度依赖性呈一级反应;(g)质粒转化效率较低,每个转化体需要摄取10³至10⁴个DNA分子。基于这些信息,提出了一个关于染色体DNA、质粒DNA和转染DNA处理过程的模型。

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