Abstract
Bacillus subtilis has tremendous applications in both academic research and industrial production. However, molecular cloning and transformation of B. subtilis are not as easy as those of Escherichia coli. Here we developed a simple protocol based on super-competent cells prepared from the recombinant B. subtilis strain SCK6 and multimeric plasmids generated by prolonged overlap extension-PCR. Super-competent B. subtilis SCK6 cells were prepared by overexpression of the competence master regulator ComK that was induced by adding xylose. This new protocol is simple (e.g., restriction enzyme, phosphatase, and ligase free), fast, and highly efficient (i.e., ~107 or ~104 transformants per μg of multimeric plasmid or ligated plasmid DNA, respectively). Shuttle vectors for E. coli–B. subtilis are not required.
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Acknowledgement
This work was supported by the DOE BioEnergy Science Center and the College of Agriculture and Life Sciences Biodesign and Bioprocessing Research Center at Virginia Tech to Y.P.Z. X.Z.Z. appreciates the support from NSF and DOE SBIR grants.
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Zhang, XZ., You, C., Zhang, YH.P. (2014). Transformation of Bacillus subtilis . In: Sun, L., Shou, W. (eds) Engineering and Analyzing Multicellular Systems. Methods in Molecular Biology, vol 1151. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0554-6_7
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DOI: https://doi.org/10.1007/978-1-4939-0554-6_7
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