Author

Lisa Kozinski

Publication Date

1-1-1998

Document Type

Dissertation/Thesis

First Advisor

Mitchell, John L. A.

Degree Name

B.S. (Bachelor of Science)

Department

Department of Biological Sciences

Abstract

The polyamines, spermidine and spermine, and their precursor, putrescine are cations that are absolutely necessary for cell growth and viability. A protein called ornithine decarboxylase antizyme was previously found to be a regulator of both polyamine biosynthesis and transport in mammalian cells. Although there is only one gene for antizyme, two forms of the protein (29.5 kDa and 24 kDa) are known to coexist. Some researchers speculate that expression of the shorter peptide (24 kDa) results from initiation of translation at a second start downstream from the primary AUG. An alternative hypothesis is that a post-translational modification of the 29.5 kDa species is necessary for expression of the 24 kDa protein. To clarify the matter, a modified form of antizyme lacking the second AUG and containing a 3’ six-histidine tag has been cloned into a eukaryotic expression vector (PMAMneo). The construct will be transfected into Chinese hamster ovary cells, and expression of the modified antizyme induced with dexamethasone. If an immunochemical screen, using antibody to the six-histidine tag, identifies a 24 kDa induced antizyme, it can be concluded that expression of the shorter protein was mediated by a post-translational modification.

Comments

Includes bibliographical references.

Extent

9 unnumbered pages

Language

eng

Publisher

Northern Illinois University

Rights Statement

In Copyright

Rights Statement 2

NIU theses are protected by copyright. They may be viewed from Huskie Commons for any purpose, but reproduction or distribution in any format is prohibited without the written permission of the authors.

Media Type

Text

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