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    Stable Maintenance of Multiple Plasmids in E. coli Using a Single Selective Marker

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    Author
    Schmidt, Calvin M.; Shis, David L.; Nguyen-Huu, Truong D.; Bennett, Matthew R.
    Date
    2012
    Abstract
    Plasmid-based genetic systems in Escherichia coli are a staple of synthetic biology. However, the use of plasmids imposes limitations on the size of synthetic gene circuits and the ease with which they can be placed into bacterial hosts. For instance, unique selective markers must be used for each plasmid to ensure their maintenance in the host. These selective markers are most often genes encoding resistance to antibiotics such as ampicillin or kanamycin. However, the simultaneous use of multiple antibiotics to retain different plasmids can place undue stress on the host and increase the cost of growth media. To address this problem, we have developed a method for stably transforming three different plasmids in E. coli using a single antibiotic selective marker. To do this, we first examined two different systems with which two plasmids may be maintained. These systems make use of either T7 RNA polymerase-specific regulation of the resistance gene or split antibiotic resistance enzymes encoded on separate plasmids. Finally, we combined the two methods to create a system with which three plasmids can be transformed and stably maintained using a single selective marker. This work shows that large-scale plasmid-based synthetic gene circuits need not be limited by the use of multiple antibiotic resistance genes.
    Citation
    Schmidt, Calvin M., Shis, David L., Nguyen-Huu, Truong D., et al.. "Stable Maintenance of Multiple Plasmids in E. coli Using a Single Selective Marker." ACS Synthetic Biology, 1, no. 445-450 (2012) American Chemical Society: http://dx.doi.org/10.1021/sb3000589.
    Published Version
    http://dx.doi.org/10.1021/sb3000589
    Type
    Journal article
    Publisher
    American Chemical Society
    Citable link to this page
    https://hdl.handle.net/1911/69864
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    • Biochemistry and Cell Biology Publications [118]

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    Home | FAQ | Contact Us | Privacy Notice | Accessibility Statement
    Managed by the Digital Scholarship Services at Fondren Library, Rice University
    Physical Address: 6100 Main Street, Houston, Texas 77005
    Mailing Address: MS-44, P.O.BOX 1892, Houston, Texas 77251-1892
    Site Map