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Naturally-Occurring Fusion Between the Regulatory and Catalytic Components of Type IIP Restriction-Modification Systems

Liang, Jixiao
http://rave.ohiolink.edu/etdc/view?acc_num=mco1376389983

Abstract Details

2013, Master of Science in Biomedical Sciences (MSBS), University of Toledo, College of Medicine.
Restriction-modification (R-M) systems play key roles in controlling gene flow among bacteria and archaea, and their own genetic mobility depends critically on their regulation, but the regulation of these systems is poorly understood. The PvuII R-M system is a Type IIP R-M system in that the protective DNA methyltransferase (MTase) is a separate and independently-active protein from the potentially lethal restriction endonuclease (REase). PvuII is one of the best studied of the R-M systems that use a positive feedback regulatory loop, involving a transcriptional regulator called C protein, to delay expression of the REase relative to that of the MTase. This allows protective methylation of a new host cell's DNA before the REase is produced. In searching for R-M systems related to PvuII, in order to study evolution and variation of its regulatory system, a putative system was found in the genome sequence of the bacterium Niabella soli strain DSM 19437, in which the regulatory C protein and the REase are translationally fused. The hypothesis is that N. soli truly produces a fused C-R protein, and that it is active as both a REase and as an autogenous regulator. The genes for the N. soli R-M system were synthesized, produced and purified with affinity tags, and the production of full-length C-REase fusion protein was confirmed. The dual activity of the fusion protein was determined by in vitro restriction of known DNAs, and in vivo transcriptional activation of a lacZ fusion to the promoter on which the C protein acts.
Robert Blumenthal, Ph.D. (Committee Chair)
Jason Huntley, Ph.D. (Committee Member)
Steve Patrick, Ph.D. (Committee Member)
58 p.
Biomedical Research
restriction-modification systems; fusion protein

Recommended Citations

Citations

  • Liang, J. (2013). Naturally-Occurring Fusion Between the Regulatory and Catalytic Components of Type IIP Restriction-Modification Systems [Master's thesis, University of Toledo]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=mco1376389983

    APA Style (7th edition)

  • Liang, Jixiao. Naturally-Occurring Fusion Between the Regulatory and Catalytic Components of Type IIP Restriction-Modification Systems. 2013. University of Toledo, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=mco1376389983.

    MLA Style (8th edition)

  • Liang, Jixiao. "Naturally-Occurring Fusion Between the Regulatory and Catalytic Components of Type IIP Restriction-Modification Systems." Master's thesis, University of Toledo, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=mco1376389983

    Chicago Manual of Style (17th edition)

mco1376389983
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© 2013, all rights reserved.
This open access ETD is published by University of Toledo Health Science Campus and OhioLINK.