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Characterizatin of a Type II Metallothionein from Helianthus annuus Using Recombinant DNA Techniques

Bhogavalli, Sridhar
http://rave.ohiolink.edu/etdc/view?acc_num=csu1198989836

Abstract Details

2007, Master of Science in Chemistry, Cleveland State University, College of Science.
Metallothioneins (MTs) are ubiquitous low molecular weight, cysteine rich proteins with a pronounced affinity for metal ions with d10 configuration such as Cu, Cd and Zn. These heavy metals form metal-thiolate clusters with cysteine side chains. In contrast to the vertebrate forms, knowledge about the properties of members of the plant metallothionein family is still scarce. We describe here a method of isolation and purification of metallothionein from the plant Helianthus annuus, performed by affinity chromatography using glutathione-agarose column. In this study, alignment of gene sequence of the isolated H. annuus metallothionein cDNA with known MTs showed that it belongs to the type 2 of the plant MTs. The gene sequence encoding MTs was cloned into a suitable vector and the protein was overexpressed in Escherichia coli. The purified metallothionein was evaluated by SDS-PAGE and characterized by UV spectra of the apo-and metal bound protein. Detection of metal bound protein was also carried out by using chemiluminescence assay. The metal binding ability of metallothionein was evaluated by 5, 5 dithio (2-nitrobenzoic acid) (DTNB) analysis. The affinity of metal ions for metallothionein was in the order of Cu>Cd>Zn.
Robert Wei (Advisor)
Lily Ng (Other)
Harry Van Keulen (Other)
45 p.
Biology, Molecular; Chemistry, Analytical
proteins; METALLOTHIONEIN; metal; MT; Fusion proteins; Gel; GST-MT

Recommended Citations

Citations

  • Bhogavalli, S. (2007). Characterizatin of a Type II Metallothionein from Helianthus annuus Using Recombinant DNA Techniques [Master's thesis, Cleveland State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=csu1198989836

    APA Style (7th edition)

  • Bhogavalli, Sridhar. Characterizatin of a Type II Metallothionein from Helianthus annuus Using Recombinant DNA Techniques. 2007. Cleveland State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=csu1198989836.

    MLA Style (8th edition)

  • Bhogavalli, Sridhar. "Characterizatin of a Type II Metallothionein from Helianthus annuus Using Recombinant DNA Techniques." Master's thesis, Cleveland State University, 2007. http://rave.ohiolink.edu/etdc/view?acc_num=csu1198989836

    Chicago Manual of Style (17th edition)

csu1198989836
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© 2007, all rights reserved.
This open access ETD is published by Cleveland State University and OhioLINK.