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Date of Award

2004

Document Type

Thesis - Pacific Access Restricted

Degree Name

Master of Science (M.S.)

Department

Biological Sciences

First Advisor

Craig A. Vierra

First Committee Member

Geoff Lin-Cereghino

Second Committee Member

Lisa A. Wrischnik

Abstract

Activated B-cell Factor-I (ABF-1) is a class II basic helix-loop-helix protein expressed in activated B-cells, EBV -immortalized lymphoblastoid cell lines and embryonic skeletal muscle in mice. ABF -1 dimerizes with another bHLH protein E4 7 to form a transcriptional repressor ofE2A, which is a gene essential to the proper development of B- and T -cells. In an effort to study genes in B-cells regulated by ABF- 1, I have attempted to construct a Burkitt Lymphoma cell line allowing tetracycline regulated expression of ABF-1. The tetracycline repressor gene was first added to these cells, creating a parental line oftet repression. Next, both a full-length and a truncated version of ABF -1 were added to the cell line. The truncated version of the protein is expressed in the presence of tetracycline, but completely repressed in its absence, demonstrating a tightly-regulated system of inducible expression. The full-length version of ABF-1 has yet to be expressed in this cell line; however, it has been expressed in a HeLa cell line, demonstrating that the construct has been properly made.

Pages

37

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