• Login
    View Item 
    •   Carroll Scholars Home
    • Life and Environmental Sciences
    • Life and Environmental Sciences Undergraduate Theses
    • View Item
    •   Carroll Scholars Home
    • Life and Environmental Sciences
    • Life and Environmental Sciences Undergraduate Theses
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Development Of A Multifunctional Escherichia coli/Bacillus subtilis Shuttle Expression Plasmid With Synthetic Expression Control

    Thumbnail
    View/Open
    1987_LehmanT_THS_000927.pdf (2.226Mb)
    Author
    Lehman, Thomas
    Advisor
    James Manion; John Addis; Richard Lambert
    Date of Issue
    1987-04-01
    Metadata
    Show full item record
    URI
    https://scholars.carroll.edu/handle/20.500.12647/3101
    Title
    Development Of A Multifunctional Escherichia coli/Bacillus subtilis Shuttle Expression Plasmid With Synthetic Expression Control
    Type
    thesis
    Abstract
    To simplify the screening of cloned DNA and to enhance the expression of protein products, a plasmid, pSP 10, was designed and constructed containing several features. These features included: 1) an origin of replication for Escherichia coli and Bacillus subtilis. 2) a selection gene, 3) an indicator gene, 4) a cloning region, and 5) a region of expression control. The plasmid shuttle vector, pSP 10, contains a synthetic promoter sequence, which was used to direct the expression of the Lac Z gene. The Lac Z gene codes for the protein B-Galactosidase. Two secretion leader sequences were also constructed and cloned into the plasmid pUC 19 for characterization. They were labelled "PEN I" and "PEN II". When properly inserted upstream of (read before) a protein-coding sequence, the secretion leader would direct secretion of the translated protein across a cell membrane in either E. coli or EL gubfilig. The project involved growing the cells and purifying the pSP 10 plasmid DNA, analyzing the DNA for the synthetic promoter sequence, and selecting restriction sites in pSP 10 which would cut the plasmid to accept the secretion leader sequence. Analysis of the plasmids, pUC/PEN I and pUC/PEN II, by restriction endonuclease mapping resulted in the identification of restriction sites compatible with cloning PEN II into pSP 10.
    Degree Awarded
    Bachelor's
    Semester
    Spring
    Department
    Life & Environmental Sciences
    Collections
    • Life and Environmental Sciences Undergraduate Theses

    Browse

    All of Carroll ScholarsCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

    My Account

    LoginRegister

    DSpace software copyright © 2002-2023  DuraSpace
    DSpace Express is a service operated by 
    Atmire NV