Date of Award

1-1-2012

Language

English

Document Type

Master's Thesis

Degree Name

Master of Science (MS)

College/School/Department

Department of Biological Sciences

Content Description

1 online resource (viii, 99 pages) : PDF file, illustrations (some color)

Dissertation/Thesis Chair

Marlene Belfort

Committee Members

Ing Nang Wang, David Shub

Keywords

Tuberculosis, Multidrug-resistant tuberculosis, Mycobacterium tuberculosis

Subject Categories

Biology

Abstract

Tuberculosis, caused by Mycobacterium tuberculosis, is one of the most widespread and highly contagious diseases in the world today. This disease affects people from all walks of life and has a devastating effect on immune compromised individuals. Another concerning fact is the emergence of multiple drug resistant and extreme drug resistant strains of the bacilli that has made treatment very difficult. Thus it has become imperative that every avenue should be explored to fight and eradicate this disease. One such unexplored avenue is the role of inteins in mycobacterial biology. Inteins are intervening sequences found in conserved regions of a gene. They splice out autocatalytically in a post-translational event only after which the host protein can be functional. Inteins have been discovered in three genes in M. tuberculosis, recA, sufB and dnaB that are critical for various cellular functions. Study of intein splicing will allow insight on whether these inteins have any regulatory role and if they can be used as potential drug targets. A toolkit needed to be developed that would allow study of intein splicing in situ. In this study a reporter system was developed by making GFP::intein constructs where fluorescence would be expressed only after the intein spliced out. Also this construct could be used for Western blotting which provides another way of monitoring splicing activity. The results of this project show that such a construct can indeed be used to study intein splicing in vivo as well as in vitro.

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