The Process of Isolating, Purifying, And Analyzing the GFP Gene from the pGLO Plasmid, and Transforming and Expressing GFP in E. coli Cells.

The aim of this project was to isolate, amplify, and purify the GFP gene from the pGLO plasmid, for both analysis as well as the translation and expression into Escherichia coli cell cultures. The Polymerase Chain Reaction (PCR) technique was used to isolate and amplify the GFP gene, which was then purified for use in the agarose gel electrophoresis (AGE) chamber, to distinguish the DNA fragments by size and determine the presence of the desired amplicon. The NanoDrop, SnapGene, and Basic Local Alignment Search Tool (BLAST) softwares were then used for the analysis of the purified amplicon. During the analysis of the purified amplicon, the A260/A280 value was 1.94. The pGLO plasmid was then transformed into competent E. coli cells, which were then left to incubate and grow for final analysis of the gene. The pGLO plasmid was successfully transformed into the E. coli cells. There was growth in all of the agarose gel plates except for the -pGLO LB/amp plate. The GFP gene from the pGLO plasmid was successfully expressed only in the cell cultures of the +pGLO LB/amp/ara agarose gel plate, as that plate had the arabinose in the agarose, which was necessary for the growth and development of the GFP gel\u27s expression within the bacteria