Abstract
Large molecular weight plasmids are often used in gene therapy and DNA vaccines. To investigate the effect of plasmid size on the performance of Escherichia coli host strains during plasmid preparation, we employed E. coli JM109 and TOP10 cells to prepare four plasmids ranging from 4.7 to 16.8 kb in size. Each plasmid was extracted from JM109 and TOP10 cells using an alkaline lysis mini-preparation method. However, when commercial kits were used to extract the same plasmids from JM109 cells, the large molecular weight plasmids substantially degraded, compared with their smaller counterparts. No degradation was observed when the four plasmids were extracted from E. coli TOP10 cells using the same commercial kit. We conclude, therefore, that the performance of E. coli in high quality plasmid preparations can be affected by plasmid size.
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Acknowledgments
This work was supported by the Natural Science Foundation of Zhejiang Province in China (Grant No. Y2111122), and the Medical and Health Research Program of Hangzhou in China (Grant No. 20110833B33).
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Yang, J., Yang, Y. Plasmid size can affect the ability of Escherichia coli to produce high-quality plasmids. Biotechnol Lett 34, 2017–2022 (2012). https://doi.org/10.1007/s10529-012-0994-4
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DOI: https://doi.org/10.1007/s10529-012-0994-4