PGlo Transformation Lab

Genetics – pGlo Transformation Lab Report
Your Name and Lab Members Names
Your Subject and Section
Introduction
In order to understand the objective of this pGlo Transformation lab, the author of this article believes that it would be best if the idea of DNA and its constituents would first be discussed. As we learned in Unit three, DNA is naturally found as a twofold helix, implying that it has two strands and is produced from nucleotides with each and every one of it nucleotide comprised of a deoxyribose sugar that is fortified by a phosphodiester grasp to a phosphate mixture. Aside from this, the nucleotide is also attached to a nitrogen base, which is held firmly by a glycosidic bond. This nitrogen base will coordinate to its correlative nitrogen base on the other strand. However, as the process of DNA replication happens, these hydrogen bonds preserving the two strands together are then damaged by way of the Helicase protein. This process would continue on until it reaches the point where the DNA becomes replicated with either a new segment or a completely similar one depending on the external factors present. In line with this, the author believes that the two exceptional subjects that we should be familiar with are the Operon system and Biotechnology. Based on our previous discussions, we have learned that the Operon system can both be repressible or inducible. Knowing this idea, scientists and other practitioners of biotechnology have utilized this in order to create different traits by mixing different portions of genetic materials using laboratory conditions. The two most common ways of doing this in a laboratory are (1) calcium chloride shock and/or (2) electroporation. In the history of biotechnology, it was Frederick Griffith who was considered as the father of modern genetics since he was the very first person to change the genetic material and the traits of an organism under lab conditions back in 1928 CITATION NCBnd \l 1033 (O'Connor, 2008).
In relation to this experiment, the researchers have utilized Escherichia coli or E. coli because of the relative ease of changing its genetic material as well as its practicality and safeness of use. More particularly, what made this one of the most chosen candidates for this experiment is because it is safe and could easily be killed by Ampicillin (antibiotic with beta-lactam structure), to act as a “safety breaker” in any case that an accident happens CITATION Kon10 \l 1033 (Kong, Schneper, & Mathee, 2010).
Aside from the two terms stated above, another concept that would be very helpful to consider are the plasmids. These circular structure would be the focus of our experiment since sections of these could easily be altered with other genetic material so that we can replace and get different resulting genetic structures. Therefore, this also means that protein production could easily be altered as we modify the genetic structures of these plasmids. However, since plasmids also vary depending on their genetic structure, the researchers have specifically chosen the pGLO plasmid because it provides an easy access for the proteins and enzymes that we would need, which are the 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. As for our analysis, the author of this article would utilize UV light (as discussed in the latter sections) to be able to see if the E Coli. Would be able to produce the GFP in different conditions. The different conditions would also be discussed further on the chapters of materials and methods.
Materials and Methods
As stated earlier, the main objective objective of the researchers in this experiment is to be able to find out if certain conditions would promote the genetic replication of E. Coli and the expression of a trait by adding a gene that is originally foreign from its genetic code. In order to do this, one of the methods employed is by adding Calcium Chloride to different solutions with one of them having an antibiotic while the others don’t. Following from the initial instructions presented, the researchers created a control setup as well as variable ones, which contains different concentrations of particular materials and genetic codes. In one of the petri dishes taken, they have replaced Calcium chloride with Distilled water to check what would be the effect on the bacterial growth as well as the process of DNA replication. All of these materials could be seen in the Appendix 1, while the setup of the petri dishes are also exhibited in Appendix 2. All in all, we were able to create a total of 4 petri dishes. Although the concentration and presence of the materials differs from each and every one of them, the researchers have added agar solutions to every one of them to be able to provide the necessary nutrients for bacterial growth, and to eliminate (lack of food source) it as an affecting variable. Some of these additional solutions are LB broth, ampicillin, as well as the arabinose enzyme. As could be seen in Appendix 2, the petri dishes were labelled as follows: (1) LBA (-pGlo), (2) LB (-pGLo), (3) LB (amp and ara), and (4) LB (+pGlo). The two plates (LB/amp) and (LB/am...