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Conjugation Mediated Bacterial Genome Mixing (CoMBacGeMi) to transfer and detect genes of interest in Escherichia coli and unravel the recombination patterns

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dc.contributor.advisor Tenaillon, Olivier
dc.contributor.author CHAVAN, ARSH
dc.date.accessioned 2023-05-19T05:29:40Z
dc.date.available 2023-05-19T05:29:40Z
dc.date.issued 2023-04
dc.identifier.citation 50 en_US
dc.identifier.uri http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/7925
dc.description.abstract Conjugation Mediated Bacterial Genome Mixing (CoMBacGeMi) is a technique developed to enable the exchange of genetic material between bacterial strains, in a manner that simulates the process occurring in the wild. CoMBacGeMi aims to elucidate the molecular basis of functional diversity observed in medically significant bacterial species, such as Escherichia coli. The method employs a library of strains, each harboring a conjugative plasmid integrated at a random position in the genome. In this study, we employed CoMBacGeMi to investigate the effect of a mutant matP recipient on the bias in recombination frequency observed at the terminus. Analysis of the pools revealed that MatP does not significantly regulate the recombination pattern. To go beyond a single locus and to investigate the recombination pattern across the entire genome, we utilized the Hfr library and the recipient tse2 library. Our results revealed that the recombination frequencies across the genome were non-uniform, with regions such as the Ter macrodomain exhibiting low recombination frequencies. Additionally, we observed that the conjugation efficiencies of recipient clones were related to the position of the loci of selection in the genome. Lastly, we tested the ability of CoMBacGeMi to identify specific targets of selection. Analysis of the pools enabled us to identify, with precision, the three distinct insertion positions of tse2 in the three tested clones. Our results demonstrate that CoMBacGeMi is a simple, precise, and accurate method with the potential to identify key genetic loci and provide insight into the complexities of genetic traits in diverse bacterial populations. en_US
dc.description.sponsorship INSERM Dr Paris IDF Centre Nord DST INSPIRE fellowship en_US
dc.language.iso en en_US
dc.subject Conjugation en_US
dc.subject Recombination en_US
dc.subject Genome Mixing en_US
dc.subject E. coli en_US
dc.subject matP en_US
dc.title Conjugation Mediated Bacterial Genome Mixing (CoMBacGeMi) to transfer and detect genes of interest in Escherichia coli and unravel the recombination patterns en_US
dc.type Thesis en_US
dc.description.embargo One Year en_US
dc.type.degree BS-MS en_US
dc.contributor.department Dept. of Biology en_US
dc.contributor.registration 20181034 en_US


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  • MS THESES [1705]
    Thesis submitted to IISER Pune in partial fulfilment of the requirements for the BS-MS Dual Degree Programme/MSc. Programme/MS-Exit Programme

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