pGLO™ Transformation and Purification of Green Fluorescent Protein (GFP)/pGLO质粒转化及绿色荧光蛋白的纯化实验

Cell transformation is the genetic alteration of a cell resulting from the direct uptake and incorporation of exogenous genetic material through cell membrane.

Uptake of foreign DNA plasmid

While all the chromosome contains large and essential information, a plasmid is a small DNA molecule within a cell that is physically separated from a chromosomal DNA and can replicate independently.

Part of the plasmid can be modified to join with a complete DNA strand and produced protein of interest. In this experiment, an engineered plasmid, pGLO, is transformed into E.Coli. In biotechnology, the rDNA (recombinant DNA) generates the green fluorescent protein (GFP) and the ampicillin resistance gene.

About GFP:

Green Fluorescent Protein and Beta-lactamase are two twin protein product synthesised after a normal E.Coli is transformed into rDNA including pGLO. GFP is extracted from jellyfish named Aequorea victoria that glowers in the dark when given radiation of UV light. Therefore, the visual expression of GFP glowering proves the existence the successful synthesis of Beta-lactamase, which possess massive pharmaceutical and research value.

Procedure:

Day 2

Day 3

Heat-Shock procedure:

Microbiology requires specific condition for certain process to happen. In this case, the pGLO plasmid doesn't genuinely happen without "heat-Shock" procedure"