Full-length single-stranded PCR product mediated chromosomal integration in intact Bacillus subtilis.

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.

Journal of microbiological methods 2013 Mar

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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. Copyright © 2012 Elsevier B.V. All rights reserved.

Citation

Sai Wen, Jianguo Yang, Tianwei Tan. Full-length single-stranded PCR product mediated chromosomal integration in intact Bacillus subtilis. Journal of microbiological methods. 2013 Mar;92(3):273-7