Doctoral Student at the
Rex Richards Building, South Parks Road, Oxford OX1 3QU
Description of project: I was a part of Synthetic Biology team and my project involved developing genetic tool for industrial bacteria, Clostridium acetobutylicum. Various efforts have been made to develop genetic tools in order to transfer large antibiotic gene clusters in industrially important clostridial species. For this purpose, pJH71 shuttle vector was developed in Dr. Heap’s lab based on the pMTL80000 modular system. This vector consists of two replicons; ColE1 (Gram negative replicon) and pSOL1 (Gram positive replicon). The source of pSOL1 replicon is pSOL1 megaplasmid found in C. acetobutylicum ATCC 824. pSOL1 megaplasmid is 192 kilobases (kb) in size and harbours genes necessary for the production of industrially important solvents. The ability of pSOL1 replicon to replicate 192 kb megaplasmid makes it a good candidate for facilitating the cloning of large biosynthetic gene clusters which can be over 100 kb. Our main aim of the project was to investigate the effects of varying insert sizes on the pSOL1 replicon properties i.e. transformation frequency and segregational stability.
2014 - 2015 Synthetic Biochemistry department, GlaxoSmithKline
Description of project: I was a part of Synthetic Biochemistry team. My project was based on engineering laccase for the drug development process using molecular Biology methods. I was involved in expressing Trametes versicolor laccases in Pichia pastoris and Saccharomyces cerevisiae in order to replicate the literature levels of expression, to investigate the interference of His tags during the expression, to compare the expression levels using various promoters (AOX, GAL1 and ADH2) and to test the activity profile of this enzyme with various compounds.
Furthermore, my side project involved human G2A receptor and rat G2A receptor yeast antagonist assays. The G2A receptors are believed to be involved in the pathogenesis of Multiple sclerosis. I was involved in high throughput screening of compounds that would inhibit these receptors, to find a possible cure for Multiple Sclerosis.
In addition, I also got an opportunity to assist Chemistry PhD student with her project. I carried out various site directed mutagenesis of the enzyme and performed Michaelis Menten kinetics on the mutated enzyme. I also did some crystallography work.June 2013 - September 2013 Structural Biology lab, Imperial College London
Description of project: I was involved in expressing protein lipase in Pichia pastoris. It was fascinating to understand the structure of this protein, it’s interactions with surfactants and how different conditions such as temperature and pH affected it’s catalytic activity on an industrial level. This internship provided me with a better understanding about the basics of using Nuclear Magnetic Resonance (NMR) to obtain highly detailed data on both the structure and dynamics of proteins.