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Research to produce the protein that synthesize Lipoic Acid.
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RESEARCH TO PRODUCE THE PROTEIN THAT SYNTHESIZE LIPOIC ACID. REPORT INTRODUCTION My principle objective was to isolate the DNA encoding Lipoic Acid Synthase (lipA) in Aeropyrum pernix (a thermophilic bacterium), and to over express the protein (LipA). Lipoic acid is a vitamin required for the healthy growth of prokaryotic and eukaryotic cells. It is synthesised by LipA in the following reaction: DNA from Aeropyrum pernix was kindly provided by Dr. M. Kreik (Southampton University), which contained the required gene. The initial practical steps for the project were: 1). Amplifying lipA gene by polymerase chain reaction (pcr). 2). Separating the pcr products on an agarose gel. 3). Isolating the required gene present in the band. Subsequently, I carried out following steps 4). Capturing the gene in a TOPO plasmid. 5). Inserting the plasmid into Escherichia coli. 6). Allowing it to grow in a medium supplemented with ampicillin These steps allowed us to isolate bacterial...
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