Escherichia coli (E. coli) is one of the most widely studied and understood living organisms. It has been used for decades in furthering understanding of gene functionality and gene-protein mechanisms for prokaryotes and eukaryotes alike (Riley et al. 2006).
Prokaryotes, like E. coli, have their DNA in the form of a plasmid. A plasmid is a circular ring of DNA. The plasmid intended to transform the E. coli colonies for the purposes of this study was the pGLO plasmid. The pGLO plasmid contains three main genes: one for GFP (green fluorescent protein), one for ampicillin (an antibiotic) resistance, and one that codes for the sugar arabinose (Bassiri 2011). The process used to transform the E. coli bacteria with the pGLO plasmid was by means …show more content…
coli is a competent bacteria cell able to take up a pGLO plasmid by means of heat shock genetic transformation. To test this, colonies of E. coli mixed either with a pGLO plasmid or without a pGLO plasmid were placed on four Luria-Bertani (LB) agar nutrient plates. One plate had only LB, two had both LB and ampicillin, and one had LB, ampicillin and arabinose sugar. We hypothesized for this study that the E. coli mixed with the pGLO plasmid would take up the plasmid and would then thrive on the agar plates with ampicillin because the pGLO plasmid contains a gene for ampicillin resistance. It also follows then that the E. coli colonies without the pGLO plasmid (i.e. also without the ampicillin resistance gene) would die off due to the presence of ampicillin in the agar. Furthermore, we hypothesized that E. coli would grow on all of the plates except for the plate with no pGLO plasmid containing LB and ampicillin. Finally, we hypothesized that the E. coli with the added pGLO plasmid would glow on the plate with LB, ampicillin, and arabinose sugar because the arabinose would activate the GFP gene also found in the pGLO plasmid causing the E. coli to produce the protein. The results of this experiment are significant because the way that many genetic processes occur in E. coli parallel the genetic processes in most other organisms (Riley et al. 2006). As such, understanding how genetic transformation works in E. coli can aid in understanding the process for all living
Prokaryotes, like E. coli, have their DNA in the form of a plasmid. A plasmid is a circular ring of DNA. The plasmid intended to transform the E. coli colonies for the purposes of this study was the pGLO plasmid. The pGLO plasmid contains three main genes: one for GFP (green fluorescent protein), one for ampicillin (an antibiotic) resistance, and one that codes for the sugar arabinose (Bassiri 2011). The process used to transform the E. coli bacteria with the pGLO plasmid was by means …show more content…
coli is a competent bacteria cell able to take up a pGLO plasmid by means of heat shock genetic transformation. To test this, colonies of E. coli mixed either with a pGLO plasmid or without a pGLO plasmid were placed on four Luria-Bertani (LB) agar nutrient plates. One plate had only LB, two had both LB and ampicillin, and one had LB, ampicillin and arabinose sugar. We hypothesized for this study that the E. coli mixed with the pGLO plasmid would take up the plasmid and would then thrive on the agar plates with ampicillin because the pGLO plasmid contains a gene for ampicillin resistance. It also follows then that the E. coli colonies without the pGLO plasmid (i.e. also without the ampicillin resistance gene) would die off due to the presence of ampicillin in the agar. Furthermore, we hypothesized that E. coli would grow on all of the plates except for the plate with no pGLO plasmid containing LB and ampicillin. Finally, we hypothesized that the E. coli with the added pGLO plasmid would glow on the plate with LB, ampicillin, and arabinose sugar because the arabinose would activate the GFP gene also found in the pGLO plasmid causing the E. coli to produce the protein. The results of this experiment are significant because the way that many genetic processes occur in E. coli parallel the genetic processes in most other organisms (Riley et al. 2006). As such, understanding how genetic transformation works in E. coli can aid in understanding the process for all living