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Attempts Towards Site-directed Mutation of an Endonuclease Gene and Insertion of a Gene in a Bacteriophage Genome

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dc.contributor.advisor KAYARAT, SAIKRISHNAN
dc.contributor.author MAHAVEER, MANAS
dc.date.accessioned 2023-12-18T04:53:59Z
dc.date.available 2023-12-18T04:53:59Z
dc.date.issued 2023-12
dc.identifier.citation 35 en_US
dc.identifier.uri http://dr.iiserpune.ac.in:8080/xmlui/handle/123456789/8330
dc.description.abstract Anti-microbial resistance is a growing issue for the healthcare sector, with more and more pathogenic strains propping up every day. It isn’t enough to just kill the cells, as the DNA released into the environment can still confer the resistance to neighbouring cells through Horizontal Gene Transfer (HGT), and hence there is the need to develop measures that can do a two-fold job: kill the resistant-strain cells and prevent further propagation. SauUSI is a Type-IV Restriction Endonuclease that has been shown to have a “shredding” activity in methylated dsDNA. It creates several cuts between recognition sites, essentially putting the DNA through a biochemical paper-shredder. In my thesis, I aimed to engineer a lytic phage delivery system for SauUSI. The phage is responsible for cell lysis while the SauUSI is meant to shred all of the cellular DNA, thus making it meaningless in the context of HGT. Using Gibson Assembly as a synthesis method, and subsequent phage rebooting, I tried to create these phage-SauUSI constructs. The second half of the thesis focussed on identifying potential residues responsible for the specific recognition sequence of SauUSI, and generating mutations in the protein to allow us to alter it. This would allow for a widening in the range of DNA that the construct can shred and have potential therapeutic applications. en_US
dc.language.iso en en_US
dc.subject Bacteriophage en_US
dc.subject Endonuclease en_US
dc.title Attempts Towards Site-directed Mutation of an Endonuclease Gene and Insertion of a Gene in a Bacteriophage Genome 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 20181145 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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