Over the past unit of Genetics and Biotechnology, my AP Bio class and I have done various activities that have sparked my interest in genetics. One significant activity that we did was performing a genetic transformation lab using E. coli bacteria and a plasmid called pGLO. I think that this pGLO lab really gave me a taste of a hands-on work involving gene expression and tracking the progress of gene expression. The goal of the lab was to get the E. coli bacteria to accept and express new genes and we specificially selected genes for ampicillin, an antibiotic, resistance and a gene that expressed bright bioflourescence. It is an interesting concept to try and get a certain organism to express new genes especially ones that they are not naturally given to the organism or genes that the organism is not used to. But I also thought that it was even more interesting that this transaction, this delivery of genes, happens constantly in nature. Many bacteria undergo the transfer of plasmids with even bacteria of other species to help give them more advantages to adapting to their surrounding environments.
To take a look at the general cycle of what occurred during the lab from the bacteria’s perspective, we could take a look at the pGLO plasmid.

The first stop is the origin where DNA replication occurs, causing the bacterium to create more plasmids so when the bacterium reproduces, each offspring receives a plasmid. The second stop is the bla gene which codes for beta-lactamase which digests ampicillin (an antibiotic) and gives the bacteria resistance to ampicillin. The third stop is the araC gene which produces a regulatory protein that prevents RNA polymerase from transcribing the GFP gene. Since the goal is to have GFP gene be expressed to determine if the bacteria is transformed by the bla gene (since the GFP gene shows bioflourescence and acts as a tracker), arabinose is introduced which induces the araC gene. The fourth part is the GFP gene which codes for a green fluorescent protein, GFP, that fluoresces under UV light.
Within this experiment, we had two groups, one called -pGLO (the control group which was not given a plasmid to transform) and one called +pGLO (the experimental group which had a plasmid inserted and transformed). These groups each had three respective dishes which tracked the progress of gene expression. The first column of dishes showed bacterial growth without any other conditions, the second column of dishes showed bacterial growth if ampicillin was added, and the third column showed bacterial growth if ampicillin was added and whether the GFP gene was expressed.

The results showed that the +pGLO plate with the inserted plasmid was able to successfully express the genes for ampicillin resistance and GFP.
And as I mentioned before, the hands-on work of being able to implement genes was very cool because there was technology and different bacteria that I got to handle and work with. For example, in this lab we heavily used pipettes and I felt like I greatly improved in my skills of using them accurately. In addition, it was interesting to handle the bacteria and use different plates to promote growth and glow and use inoculating loops to transfer colonies to certain plates.
Overall, this pGLO lab was very interesting and gave me new insight into the revelation of genetic transformation and how this concept can be applied to living organisms.
































