Degree Type

Dissertation

Date of Award

1995

Degree Name

Doctor of Philosophy

Department

Veterinary Microbiology and Preventive Medicine

First Advisor

Robert E. Andrews, Jr.

Abstract

Mycobacterium paratuberculosis transcriptional and translational signals were studied to gain insight into gene expression in this organism. To study M. paratuberculosis promoter structures, a more versatile promoter selection vector was constructed from the pKO1 parent vector. This new plasmid, pJJ2, was used to identify 11 promoter fragments from an M. paratuberculosis DNA library. In addition, a previously characterized M. paratuberculosis promoter, contained in a 493-bp EcoRI fragment, was cloned into the new vector to test the efficacy in cloning novel M. paratuberculosis promoters. In a related study, an expression probe shuttle plasmid (pYUB76) was employed to clone M. paratuberculosis expression signals that could be studied directly in mycobacteria. Using this vector to identify M. paratuberculosis expression signals, we have determined the nucleotide sequence of ten promoter-containing fragments and have compared these sequences to those of several previously reported Mycobacterium promoters. Hexanucleotide sequences centered approximately 35 and 10 base pairs upstream from the experimentally determined transcription start sites revealed a consensus that is different from E. coli. Compilation of these promoter sequences identified the -35 (T, T/G, G, A/G, G, T) and the -10 (C, A, G, C, C, G) conserved hexanucleotides.

DOI

https://doi.org/10.31274/rtd-180813-10164

Publisher

Digital Repository @ Iowa State University, http://lib.dr.iastate.edu/

Copyright Owner

John Patrick Bannantine

Language

en

Proquest ID

AAI9610937

File Format

application/pdf

File Size

129 pages

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