Optimisation of transformation in the heterologous fusion gene GAPDH-IFN cloning using DH5α strain of Escherichia coli
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Authors
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Anisha Valsalam
ICAR-CIFE
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K. V. Rajendran
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Pooja Vinde
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Megha Kadam Bedekar
Abstract
DNA vaccines are the most essential tool of the disease prevention strategy. In the present study, cloning of a heterologous fusion gene involving glyceraldehyde 3-phosphate dehydrogenase and interferon gamma (GAPDH-IFN) was conceptualised. Cloning was tried using four distinct transformation techniques viz. InstAclone PCR Cloning kit (Fermentas, USA); CaCl2 transformation protocol; Clontech stellar competent cells protocol and PEG 8000-mediated transformation method, for the heterologous GAPDH-IFN fusion gene using DH5α strain of Escherichia coli. The first three methods were found to be unsuitable, and the PEG 8000-mediated transformation method yielded positive clones.
Keywords: DNA vcaccine, Edwardsiella tarda, Fusion genes, Labeo rohita, PEG 8000, Transformation methods
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