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Title: Functional Characterization of Recombinant and#945; Crystallin of Mycobacterium tuberculosis H37Rv
Researcher: Krishnan Gautam
Guide(s): UTPAL ROY
Keywords: Biochemistry and Molecular Biology
Biology and Biochemistry
Life Sciences
University: Birla Institute of Technology and Science
Completed Date: 2019
Abstract: Tuberculosis is an ancient disease that has reappeared in the developed world due to newlinerapid urbanization, immigration and is a major threat in the developing world due to poor newlineimmune and nutritional status. It is caused by Mycobacterium tuberculosis (M. tb). There newlineis a need to develop new vaccines and diagnostics in view of the variable efficacy of the newlineBCG vaccine in adults and the phenomenon of latent tuberculosis (TB). The HIV newlineepidemic also plays a role in suppressing the immune system; leading to reactivation of newlinethe tubercle bacilli even in the latent form. The aim of this study was to clone, express newlineand characterize important proteins associated with latent TB and immune evasion with newlinethe aim of generating new data for important biomarker proteins for diagnosis. In lieu of newlinethis, attempts were made to clone and express three M. tb proteins; superoxide dismutase newlineA (Sod A), superoxide dismutase C (Sod C) and -crystallin (Acr), known to be newlineupregulated in a macrophage model developed in human monocytic cell line (THP-1). In newlinethe present study, a total of five constructs were obtained; two of sod A, one of sod C and newlinetwo of acr. While undertaking gene-cloning experiments, the success of cloning newlineexperiments depends upon a number of factors including quality of DNA, units of newlinerestriction enzymes and ligase, molar ratios of vector and insert, high efficiency of newlinecompetent cells. newlineTraditional ligation dependent cloning methods entail use of DNA modifying enzymes newlinethat are inefficient in spite of being expensive; affecting overall cloning success. newlineParameter analysis was used in Microsoft Excel to propose key observations and newlinerecommendations for optimum results. Based on the above data, a broad model for newlineprediction of cloning success was developed which included eighteen parameters. This newlinewas applied to fifteen cases out of which nine were negative and six were positive newlineoutcomes. In addition, we used different classifiers in MATLAB and Clone. R to predict newlinevii newlinepossibility of obtaining a clone. In MATLAB, the Simple Tree classifier
Pagination: 191p.
Appears in Departments:Biological Science

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