全长单链 PCR 产物介导的完整枯草芽孢杆菌染色体整合。
Full-length single-stranded PCR product mediated chromosomal integration in intact Bacillus subtilis.
机构信息
Beijing Key Laboratory of Bioprocess, The Biorefinery Research and Engineering Center of the Ministry of Education of China, College of Life Science and Technology, Beijing University of Chemical Technology, Beijing, 100029, PR China.
出版信息
J Microbiol Methods. 2013 Mar;92(3):273-7. doi: 10.1016/j.mimet.2012.11.012. Epub 2012 Nov 28.
The research introduced a novel method for gene replacement in intact Bacillus subtilis by employing full-length single-stranded (ss) DNA constructs and electro-transformation. 5' phosphorothioated lagging-strand targeting ssDNA construct was demonstrated to be highly recombinogenic, and the utility of the system was illustrated by introducing a heterologous lipase YlLip2 into amyE locus of B. subtilis through our method.
该研究通过使用全长单链（ss）DNA 构建体和电转化，为完整枯草芽孢杆菌中的基因替换引入了一种新方法。实验证明，5' 硫代磷酸化的滞后链靶向 ssDNA 构建体具有高度的重组能力，通过我们的方法将异源脂肪酶 YlLip2 引入枯草芽孢杆菌 amyE 基因座，证明了该系统的实用性。
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