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The Establishment and characterization of a novel plant in vitro cleavage and polyadenylation assay system

Zheng, Jun
http://rave.ohiolink.edu/etdc/view?acc_num=miami1281031132

Abstract Details

2010, Master of Science, Miami University, Botany.
Cleavage and polyadenylation are two tightly coupled reactions required for messenger RNA processing in eukaryotic cells. An in vitro biochemical assay is critical for studying this process, but lack of such an assay system in plants hampered the study of plant mRNA 3’-end formation for the last two decades. This thesis reports the establishment and characterization a plant in vitro cleavage and polyadenylation assay system, in which soluble nuclear protein extracts from Arabidopsis cell cultures can accurately cleave different pre-mRNAs at expected poly(A) sites. The cleavage reaction was performed at optimal condition of 3.5mM Mg2+ by an endoribonuclease activity, with a Km of 15.3nM. When supplemented with yeast poly(A) polymerase, about 150-nucleotide poly(A) tracts were added specifically to the newly cleaved 3’-ends. This long-sought system will serve as a platform for further studies of plant mRNA 3’-end formation mechanisms and other RNA processing events.
Qingshun Q. Li, PhD (Advisor)
Chun Liang, PhD (Committee Member)
John W. Hawes, PhD (Committee Member)
37 p.
Biochemistry; Botany
Arabidopsis; pre-mRNA; in vitro; cleavage; polyadenylation

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Citations

  • Zheng, J. (2010). The Establishment and characterization of a novel plant in vitro cleavage and polyadenylation assay system [Master's thesis, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1281031132

    APA Style (7th edition)

  • Zheng, Jun. The Establishment and characterization of a novel plant in vitro cleavage and polyadenylation assay system. 2010. Miami University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1281031132.

    MLA Style (8th edition)

  • Zheng, Jun. "The Establishment and characterization of a novel plant in vitro cleavage and polyadenylation assay system." Master's thesis, Miami University, 2010. http://rave.ohiolink.edu/etdc/view?acc_num=miami1281031132

    Chicago Manual of Style (17th edition)

miami1281031132
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© 2010, all rights reserved.
This open access ETD is published by Miami University and OhioLINK.