利用质粒DNA对枯草芽孢杆菌原生质体进行高频转化。
High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA.
作者信息
Chang S, Cohen S N
出版信息
Mol Gen Genet. 1979 Jan 5;168(1):111-5. doi: 10.1007/BF00267940.
PMID:107388
Abstract
A highly efficient method for transformation of Bacillus subtilis by plasmid DNA is reported. The procedure, which involves polyethylene glycol-induced DNA uptake by protoplasts and subsequent regeneration of the bacterial cell wall, yields up to 80% transformants with an efficiency of 4 x 10(7) transformants per microgram of supercoiled DNA. Plasmids constructed by in vitro ligation or endonuclease-generated fragments of linear plasmid DNA can also transform PEG-treated protoplasts, but at a lower frequency.
摘要
报道了一种通过质粒DNA转化枯草芽孢杆菌的高效方法。该方法包括聚乙二醇诱导原生质体摄取DNA以及随后细菌细胞壁的再生，可产生高达80%的转化体，每微克超螺旋DNA的转化效率为4×10⁷个转化体。通过体外连接构建的质粒或线性质粒DNA的核酸内切酶产生的片段也可以转化经聚乙二醇处理的原生质体，但频率较低。
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