Electroporation is a feasible method to introduce circularized or linearized DNA into <Emphasis Type="Italic">B. subtilis</Emphasis> chromosome |
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Authors: | Ming-Ming Yang Wei-Wei Zhang Xiao-Ting Bai Heng-Xin Li Pei-Lin Cen |
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Institution: | (1) College of Animal Sciences, Northwest A&F University, 712100 Yangling, People’s Republic of China;(2) Institute of Biochemical Engineering, College of Material Science and Chemical Engineering, Zhejiang University, 310027 Hangzhou, People’s Republic of China;(3) Yangguang-Guangji Medical R&D co. Lt, 430062 Wuhan, People’s Republic of China |
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Abstract: | Here, we present the electroporation as a feasible and efficient method for introducing circularized and linearized DNA into
Bacillus subtilis chromosome. Two integration experiments were carried out and demonstrated the feasibility and efficiency of electroporation
to introduce the target DNA into the B. subtilis chromosome. By using of electroporation, a multiple-cistron contained five genes from B. subtilis biotin biosynthetic pathway was introduced into the B. subtilis chromosome efficiently and created a repeated copy in chromosome via a single crossover event. Then an ectopic promoter was
introduced conveniently into the upstream of one of the repeated multiple-cistron via a double crossover event. To further
demonstrate the application of electroporation in genetic research, the early sporulation gene spo0A of B. subtilis was knocked out and, consequently, the null of sporulation and logged growth was observed in this study. Thus, the electroporation
as an alternative method of integration in B. subtilis is feasible and practical. |
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