Cline S W, Lam W L, Charlebois R L, Schalkwyk L C, Doolittle W F
Department of Biochemistry, Dalhousie University, Halifax, N.S., Canada.
Can J Microbiol. 1989 Jan;35(1):148-52. doi: 10.1139/m89-022.
We present a practical description of polyethylene glycol mediated spheroplast transformation of Halobacterium halobium and Halobacterium volcanii. This method has been applied to phage DNA transfection, plasmid DNA transformation, and transformation with linear fragments of high molecular weight genomic DNA. Efficient spheroplast regeneration allows uncomplicated recovery of transformed progeny. Transformations can be performed equally well using fresh or frozen cell preparations. These methods should find application in molecular cloning, genetic fine mapping, and strain construction.
我们介绍了聚乙二醇介导的盐生盐杆菌和火山盐杆菌原生质体转化的实用方法。该方法已应用于噬菌体DNA转染、质粒DNA转化以及高分子量基因组DNA线性片段的转化。高效的原生质体再生使得转化后代的回收变得简单。使用新鲜或冷冻的细胞制剂进行转化的效果同样良好。这些方法应可应用于分子克隆、基因精细定位和菌株构建。
