The overall goal of these experiments is to investigate the expression of Green Fluorescent Protein (GFP) in bacterial cell. In other words, indicating the insertion of GFP into the target lepA/EF-4 to create a functional fusion protein.
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–Intro/background: What GFP is? he jellyfish green fluorescent protein (GFP) can be inserted into the middle of another protein to produce a functional, fluorescent fusion protein… Why GFP? GFP was the first intrinsically fluorescent protein discovered and the first shown to be useful as a fluorescent marker for analysis of cell structure and function… What lepA/EF-4 ? All living systems employ mRNA to convert genetic information (stored in nucleic acid polymers) into the functional forms of genes (expressed usually as polymers of amino acids). The process of converting mRNA into protein is known as protein synthesis, or translation; and it is one of the most highly conserved processes that living cells carry out. Protein synthesis requires ribosomes, many protein factors, and occurs in three stages; initiation, elongation, and termination… Initially the target plasmid (pPEM109) containing the lepA gene will be isolated and donor transposon DNA will be synthesized. After characterizing these pieces of DNA, they will be combined with Tn5 transposes. Also, include the a brief overview about what we did in lab taking this info from the material/method file below.
