Chen, Jiemin (2010)
Ph.D. thesis, University of Birmingham.
Mycobacterium tuberculosis, the causative agent of the infectious disease tuberculosis, has a distinct lipid-rich cell wall. Several anti-TB drugs target cell wall biosynthetic pathways. A good understanding of its biosynthesis will provide helpful clues for the development of novel drug targets. A strategy based on random transposon (Tn) mutagenesis with two different screening criteria, altered colony morphology and mycobacteriophage resistance, was developed. Non-pathogenic Mycobacterium smegmatis and the fish pathogen Mycobacterium marinum were used for generating Tn-mutant libraries. From the colony morphology screen, two out of eight genes identified from M. smegmatis Tn-mutants and eleven out of twenty from M. marinum Tn-mutants with altered colony morphology were directly involved in cell wall synthesis. One mutant from each species was chosen for further study, the M. smegmatis 3D9 Tn-mutant with the only isocitrate dehydrogenase (icd) gene disrupted and the M. marinum 8G10 mutant with a Tn inserted into the promoter region of MMAR0978 with a deficiency in methoxymycolates production. Four mutants resistant to generalised transducing phage I3 were identified with a Tn insertion in a gene cluster involved in the biosynthesis of the cell wall associated glycopeptidolipids (GPLs), demonstrating the potential of using phage-resistant mutants for identifying cell wall biosynthetic genes.
This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder.
Repository Staff Only: item control page