The Newly Identified Proteins Involved in the Chromosome Segregation in Mycobacteria- the Functional Analysis

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Chromosome Segregation: Chromosome segregation in bacteria is an active process involving a number of proteins. In most of the bacteria, it depends on the presence of ParABS system. ParB segregation protein binds to the parS sequence and forms segregation complexes. ParA segregation protein interacts with ParB complexes being responsible for their distribution. ParA homologs: C. cresentus MipZ, C. glutamicum PldP. MipZ exhibited nonspecific DNA binding activity similar to ParA-like proteins. Characteristic Features of Mycobacteria: Rod shaped, Slow growing, Gram positive bacteria with high ( G + C ) content (61 - 71% ), Lipid rich cell wall, Resistant to many environmental factors: drying, high and low temperature, high and low pH. Unique chromosome organisation and segregation in Mycobacteria:Chromosome is positioned asymmetrically in M. smegmatis cells. oriC is in the off-centre position after oriC duplication, new pole segrosome migrates faster than old pole segrosome. Mycobacterial cell elongate asymmetrically – the old pole extends faster. ParA in M. smegmatis: ParA is responsible for the localisation of the ParB complexes in the cell. parA deletion strain is characterized by slow growth. parA deletion results in formation 30% of anucleate cells. MSMEG_3743 (Soj) role in M. smegmatis:MSMEG_3743 interacts with ParB segregation protein in bacterial two hybrid system. Deletion of msmeg_3743 gene results in: Slower growth rate, Division disorders and occurence of very long cells (over 6,6 μm), Disorders in chromosome segregation (19,8 % of anucleate cells). The aim of Msc project: Localisation of MSMEG_3743 (Soj) in wild type M. smegmatis background. Localisation of MSMEG_3743 (Soj) in parB deletion background – elucidation of ParB influence on Soj localisation. Colocalisation of MSMEG_3743 (Soj) with ParB-mNeon. Strategies: Exchange of the msmeg_3743 to msmeg_3743-dendra fusion in the native chromosomal locus. Introduction of mcherry-msmeg_3743 in the integrative plasmid


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