BIOL 230W Assignment: pGlo Bacterial Transformation

pGlo Bacterial Transformation
Your Name and Lab Members Names
Your Institution of Affiliation
October 22, 2017
Introduction
In its most general definition, bacterial transformation happens a cellular organism takes a piece of gene from an external source and adds it up to its own genome. Although this technique is already age-old, this process still opens up new possibilities for future technological innovations in many fields of studies. In the lab, bacterial transformation is usually done through two methods: (1) through calcium chloride shock and/or (2) electroporation. While the first one is done through the incorporation of plasmids inside the “chemically-competent” cells, the latter is done through the utilization of electric shock in order to make the membrane more permeable to the specific DNA targeted. As recorded in history, the very first person to this was Frederick Griffith. Back in 1928, he was able to do this process in his lab, resulting a disastrous effect of transforming a healthy strain of bacteria into a deadly one.
In this study, the authors utilized Escherichia coli or E. coli. This bacteria is a gram negative, quick reproducing, non-fatal, and single celled bacteria, which makes it a good candidate for such types of experiments. Following from this, the researchers also have a dose of Ampicillin that has the ability to destroy the E. coli cells. This antibiotic consists of what we call as a beta-lactam structure.
Another important term that is discussed in the latter sections of this report are plasmids. Mainly, plasmids are small circular pieces of DNA that has the ability to autonomously replicate. This process of plasmid replication is usually happens as they borrow polymerase from the cell in which they reside. Plasmids could easily be experimented even in normal laboratory conditions because its size is just right to be extracted and purified from bacterial cells. And, since they are responsible for protein production and other processes, plasmid replication and modification also presents a number of possibilities in genetics and other fields related to it.
In particular, the researchers have utilized the pGLO plasmid since it contains GFP (Green Florescent Protein) as well as beta-lactamase. It also has a special gene regulation system which are used by most researchers around the world to control gene expression and protein creation. In other to study this type of replication, GFP would be measured as it becomes exposed to UV light. Since GFP becomes more apparent when exposed to UV light, then any changes in its luminescence as a result of gene manipulation makes it easier to understand. In most cases GFP production is monitored by a modified arabinose operon located on the pGLO plasmid. In this mechanism, a araC, a protein, blocks RNA polymerase from binding to the Pbad promoter. This is crucial for this experiment because when arabinose is present, the araC protein promotes RNA polymerase binding, as compared to its original function of promoting it. As this process continues, transcription of the GFP protein continues, until the enzyme arabinose runs out.
Materials and Methods
In this experiment, the objective of the researchers was to determine the effects of ...