{"id":{"repo_id":"umn","oai_identifier":"oai:conservancy.umn.edu:11299/169353"},"canonical_url":"https://search.dev.ndltd.org/etd/umn/oai:conservancy.umn.edu:11299/169353","repository":{"repo_id":"umn","name":"University of Minnesota","base_url":"https://conservancy.umn.edu/server/oai/request"},"display":{"title":"Prevalence of the gene lsr2 among the genus Mycobacterium and an investigation into changes in biofilm formation when inactivating lsr2 regulated genes in Mycobacterium smegmatis","abstract":"Mycobacterium smegmatis is a non-pathogenic model organism commonly used to study the human pathogen Mycobacterium tuberculosis. M. smegmatis has a rough colony morphology and is capable of forming biofilms, however, when the DNA-regulating protein Lsr2 is inactivated the bacteria forms a smooth colony and loses biofilm formation. This study investigates the phenotypic effect on colony morphology and biofilm formation when Lsr2-regulated genes are inactivated. A mutant strain of M. smegmatis with defective lsr2 (M. smegmatis &#916;lsr2) was used as a parent strain for the study and the gene for mycocerosic acid synthase (mas) was selected for inactivation. Genes downstream of mas from MSMEG_4727 to MSMEG_4733 and MSMEG_4741were also selected for inactivation. Genes were inactivated using homologous recombination via \"suicide vector\" plasmids lacking a Mycobacterial origin of replication and carrying kanamycin resistance and PCR fragments of the target genes. M. smegmatis &#916;lsr2 was transformed by electroporation and transformants were plated and screened for kanamycin resistance. The mutant strain lacking functional lsr2 and mas (M. smegmatis &#916;lsr2&#916;mas) regained the ability to form biofilms and exhibited colony morphology similar to wild-type M. smegmatis. Inactivation of genes MSMEG_4727 to MSMEG_4733 produced similar results. Inactivation of MSMEG_4741 caused no change in the &#916;lsr2 mutant phenotype. These results indicate that mas and genes downstream are involved with the biofilm formation and colony morphology change associated with the M. smegmatis &#916;lsr2 mutant strain.","abstract_html":"Mycobacterium smegmatis is a non-pathogenic model organism commonly used to study the human pathogen Mycobacterium tuberculosis. M. smegmatis has a rough colony morphology and is capable of forming biofilms, however, when the DNA-regulating protein Lsr2 is inactivated the bacteria forms a smooth colony and loses biofilm formation. This study investigates the phenotypic effect on colony morphology and biofilm formation when Lsr2-regulated genes are inactivated. A mutant strain of M. smegmatis with defective lsr2 (M. smegmatis &amp;#916;lsr2) was used as a parent strain for the study and the gene for mycocerosic acid synthase (mas) was selected for inactivation. Genes downstream of mas from MSMEG_4727 to MSMEG_4733 and MSMEG_4741were also selected for inactivation. Genes were inactivated using homologous recombination via &quot;suicide vector&quot; plasmids lacking a Mycobacterial origin of replication and carrying kanamycin resistance and PCR fragments of the target genes. M. smegmatis &amp;#916;lsr2 was transformed by electroporation and transformants were plated and screened for kanamycin resistance. The mutant strain lacking functional lsr2 and mas (M. smegmatis &amp;#916;lsr2&amp;#916;mas) regained the ability to form biofilms and exhibited colony morphology similar to wild-type M. smegmatis. Inactivation of genes MSMEG_4727 to MSMEG_4733 produced similar results. Inactivation of MSMEG_4741 caused no change in the &amp;#916;lsr2 mutant phenotype. These results indicate that mas and genes downstream are involved with the biofilm formation and colony morphology change associated with the M. smegmatis &amp;#916;lsr2 mutant strain.","abstract_has_math":false,"creators":["Gatlin, Wayne Charles"],"institution":null,"degree_name":null,"degree_level":null,"degree_discipline":null,"degree_department":null,"school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2014,"date_issued":"2014-10","date_published":"2014-10","updated_at":"2026-07-24T05:19:56Z","subjects":["Integrated biosciences"],"languages":["en"],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/11299/169353","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:creator","label":"Author","values":["Gatlin, Wayne Charles"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2014-12-30T16:53:25Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2014-12-30T16:53:25Z"]},{"key":"dc:date.issued","label":"Date","values":["2014-10"]},{"key":"dc:type","label":"Dc Type","values":["Thesis or Dissertation"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Integrated biosciences"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/11299/169353"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description","label":"Description","values":["University of Minnesota M.S. thesis. October 2014. Major: Integrated Biosciences. Advisor: John L. Dahl. 1 computer file (PDF); v, 66 pages."]},{"key":"dc:description.abstract","label":"Abstract","values":["Mycobacterium smegmatis is a non-pathogenic model organism commonly used to study the human pathogen Mycobacterium tuberculosis. M. smegmatis has a rough colony morphology and is capable of forming biofilms, however, when the DNA-regulating protein Lsr2 is inactivated the bacteria forms a smooth colony and loses biofilm formation. This study investigates the phenotypic effect on colony morphology and biofilm formation when Lsr2-regulated genes are inactivated. A mutant strain of M. smegmatis with defective lsr2 (M. smegmatis &#916;lsr2) was used as a parent strain for the study and the gene for mycocerosic acid synthase (mas) was selected for inactivation. Genes downstream of mas from MSMEG_4727 to MSMEG_4733 and MSMEG_4741were also selected for inactivation. Genes were inactivated using homologous recombination via \"suicide vector\" plasmids lacking a Mycobacterial origin of replication and carrying kanamycin resistance and PCR fragments of the target genes. M. smegmatis &#916;lsr2 was transformed by electroporation and transformants were plated and screened for kanamycin resistance. The mutant strain lacking functional lsr2 and mas (M. smegmatis &#916;lsr2&#916;mas) regained the ability to form biofilms and exhibited colony morphology similar to wild-type M. smegmatis. Inactivation of genes MSMEG_4727 to MSMEG_4733 produced similar results. Inactivation of MSMEG_4741 caused no change in the &#916;lsr2 mutant phenotype. These results indicate that mas and genes downstream are involved with the biofilm formation and colony morphology change associated with the M. smegmatis &#916;lsr2 mutant strain."]},{"key":"dc:title","label":"Title","values":["Prevalence of the gene lsr2 among the genus Mycobacterium and an investigation into changes in biofilm formation when inactivating lsr2 regulated genes in Mycobacterium smegmatis"]}]}],"canonical_facts":{"dc:creator":["Gatlin, Wayne Charles"],"dc:date.accessioned":["2014-12-30T16:53:25Z"],"dc:date.available":["2014-12-30T16:53:25Z"],"dc:date.issued":["2014-10"],"dc:description":["University of Minnesota M.S. thesis. October 2014. Major: Integrated Biosciences. Advisor: John L. Dahl. 1 computer file (PDF); v, 66 pages."],"dc:description.abstract":["Mycobacterium smegmatis is a non-pathogenic model organism commonly used to study the human pathogen Mycobacterium tuberculosis. M. smegmatis has a rough colony morphology and is capable of forming biofilms, however, when the DNA-regulating protein Lsr2 is inactivated the bacteria forms a smooth colony and loses biofilm formation. This study investigates the phenotypic effect on colony morphology and biofilm formation when Lsr2-regulated genes are inactivated. A mutant strain of M. smegmatis with defective lsr2 (M. smegmatis &#916;lsr2) was used as a parent strain for the study and the gene for mycocerosic acid synthase (mas) was selected for inactivation. Genes downstream of mas from MSMEG_4727 to MSMEG_4733 and MSMEG_4741were also selected for inactivation. Genes were inactivated using homologous recombination via \"suicide vector\" plasmids lacking a Mycobacterial origin of replication and carrying kanamycin resistance and PCR fragments of the target genes. M. smegmatis &#916;lsr2 was transformed by electroporation and transformants were plated and screened for kanamycin resistance. The mutant strain lacking functional lsr2 and mas (M. smegmatis &#916;lsr2&#916;mas) regained the ability to form biofilms and exhibited colony morphology similar to wild-type M. smegmatis. Inactivation of genes MSMEG_4727 to MSMEG_4733 produced similar results. Inactivation of MSMEG_4741 caused no change in the &#916;lsr2 mutant phenotype. These results indicate that mas and genes downstream are involved with the biofilm formation and colony morphology change associated with the M. smegmatis &#916;lsr2 mutant strain."],"dc:identifier.uri":["http://hdl.handle.net/11299/169353"],"dc:language.iso":["en"],"dc:subject":["Integrated biosciences"],"dc:title":["Prevalence of the gene lsr2 among the genus Mycobacterium and an investigation into changes in biofilm formation when inactivating lsr2 regulated genes in Mycobacterium smegmatis"],"dc:type":["Thesis or Dissertation"]},"updated_at":"2026-07-24T05:19:56Z"}