{"id":{"repo_id":"birmingham","oai_identifier":"oai:etheses.bham.ac.uk:1140"},"canonical_url":"https://search.dev.ndltd.org/etd/birmingham/oai:etheses.bham.ac.uk:1140","repository":{"repo_id":"birmingham","name":"University of Birmingham","base_url":"https://etheses.bham.ac.uk/cgi/oai2"},"display":{"title":"Identifying biosynthetic pathways for mycobacterial cell wall components using transposon mutagenesis","abstract":"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.","abstract_html":"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.","abstract_has_math":false,"creators":["Chen, Jiemin"],"institution":"University of Birmingham","degree_name":"d_ph","degree_level":"d_ph","degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2010,"date_issued":"2010-12","date_published":"2010-12","updated_at":"2026-07-24T01:11:23Z","subjects":["QR Microbiology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":null,"outbound_label":null,"outbound_source":null},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.sponsor","label":"Sponsor","values":["other"]},{"key":"dc:creator","label":"Author","values":["Chen, Jiemin"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date","label":"Dc Date","values":["2010-12"]},{"key":"dc:date.issued","label":"Date","values":["2010-12"]},{"key":"dc:publisher.department","label":"Dc Publisher Department","values":["College of Life & Environmental Sciences","School of Biosciences"]},{"key":"dc:publisher.institution","label":"Dc Publisher Institution","values":["University of Birmingham"]},{"key":"dc:relation.isreferencedby","label":"Dc Relation Isreferencedby","values":["http://etheses.bham.ac.uk//id/eprint/1140/"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.qualificationlevel","label":"Dc Type Qualificationlevel","values":["d_ph"]},{"key":"dc:type.qualificationname","label":"Dc Type Qualificationname","values":["d_ph"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["QR Microbiology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://etheses.bham.ac.uk//id/eprint/1140/1/Chen_10_PhD.pdf","http://etheses.bham.ac.uk//id/eprint/1140/2/Decl_IS_Chen_10_PhD.pdf"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["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."]},{"key":"dc:format","label":"Dc Format","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Identifying biosynthetic pathways for mycobacterial cell wall components using transposon mutagenesis"]}]}],"canonical_facts":{"dc:contributor.sponsor":["other"],"dc:creator":["Chen, Jiemin"],"dc:date":["2010-12"],"dc:date.issued":["2010-12"],"dc:description.abstract":["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."],"dc:format":["application/pdf"],"dc:identifier.uri":["http://etheses.bham.ac.uk//id/eprint/1140/1/Chen_10_PhD.pdf","http://etheses.bham.ac.uk//id/eprint/1140/2/Decl_IS_Chen_10_PhD.pdf"],"dc:publisher.department":["College of Life & Environmental Sciences","School of Biosciences"],"dc:publisher.institution":["University of Birmingham"],"dc:relation.isreferencedby":["http://etheses.bham.ac.uk//id/eprint/1140/"],"dc:subject":["QR Microbiology"],"dc:title":["Identifying biosynthetic pathways for mycobacterial cell wall components using transposon mutagenesis"],"dc:type":["Thesis"],"dc:type.qualificationlevel":["d_ph"],"dc:type.qualificationname":["d_ph"]},"updated_at":"2026-07-24T01:11:23Z"}