Future Study

The hypothesis, E. coli samples not exposed to UV light will have a higher rate of pGAL plasmid transformation success indicated by a higher blue colony count, was accepted. The data shows that the foreign DNA (pGAL plasmid) can replicate while the organism repairs its own DNA, even though the UV radiation, and possibly the DNA repair, does hinder the transformation efficiency. Therefore, if ionizing radiation and bacterial transformation combined as a form of cancer treatment, the plasmid used would replicate autonomously, yet at a lower efficiency.
However, there are internal and external radiation therapies. External radiation therapies use UV waves to shrink and eliminate tumors, while internal radiation therapies use capsules of radioactive elements, like cesium, inserted into the body. Testing the effects of internal radiation on bacterial transformation is a possible amendment to this study.

Conclusion

The hypothesis, E. coli samples not exposed to UV light will be have a higher rates of pGAL plasmid transformation success indicated by a higher blue colony count, was supported. E. coli plated with pGAL plasmid that was not exposed to UV light had higher transformation efficiency than E. coli plated with pGAL plasmid exposed to UV light. Furthermore E. coli with pGAL plasmid introduced and plated on media with ampicillin had more growth than E. coli without pGAL plasmid introduced and plated on ampicillin. Plates with ampicillin in the media inoculated with E. coli that had not been transformed with the pGAL plasmid showed no growth, indicating that the pGAL plasmid successfully codes for ampicillin resistance. Plates without ampicillin in the media inoculated with E. coli that had not been transformed by the pGAL (Control) all showed growth indicating that the E. coli was alive but did not die with radiation treatment indicating the treatment damaged the E. coli but did not kill it, as intended.
To ensure that chemicals introduced to E. coli for the successful transformation of the pGAL plasmid did not, in fact, hinder bacterial growth, and thereby skewing data, K12 E. coli was plated with each of the transformation chemicals provided. Throughout this series of testing it was found that transformation solutions provided for the pGAL plasmid transformation had no hindering effects on the growth of E. coli. Thus, solutions introduced had no adversary effects on data collected.

Hypothesis

H0 – There will be no change in rate of pGAL plasmid transformation success in E. coli samples with UV exposure compared to those with no UV exposure.
H1 – E. coli samples not exposed to UV light will have a higher rate of pGAL plasmid transformation success indicated by a higher blue colony count.

Significance

Bacterial transformation is a technique used in modern molecular biology where a foreign plasmid is introduced into bacteria causing an increased production of the protein for which the plasmid is coded. The Beta Galactosidase protein in E. coli is one encoded by the lacZ gene and is an enzyme that catalyzes the breakdown of lactose into glucose and galactase – the glucose being used as an energy source. The pGAL plasmid to be introduced contains the active lacZ gene to be coded by the E. coli. A blue color change in the E. coli colonies indicates the success of the transformation process and thus the production of Beta Galactosidase. Ultraviolet radiation therapy is used in the treatment of certain cancers and tumors. Bacterial transformation is used to produce either treatments for disease, such as insulin, or vaccines against disease. If these two processes were to be used in combination it is significant to know the effects UV radiation has on the transformation of E. coli by plasmid DNA. Furthermore, E. coli transformation may be used in the near future in treatment of cancers, and other diseases, and potentially in combination with UV radiation and other ionizing radiation therapies. Therefore this experiment, in combining the two, holds extreme medical significance to be applied in the near future.

Purpose

The purpose of this experiment is to determine the effect UV light has on the transformation process in E. coli by using a pGAL plasmid DNA sample to identify the performance of the Beta Galactosidase protein.

Abstract

The purpose of this experiment is to determine the effect UV light has on the transformation process in E. coli by using a pGAL plasmid DNA sample to identify the performance of the Beta Galactosidase protein. Bacterial transformation is a technique used where a foreign plasmid is introduced into bacteria causing an increased production of the protein for which the plasmid is coded. This new science has the potential to be used in treatments for cancer. Ultraviolet radiation is used in cancer treatments and therapies, thus it is significant to know the effects UV light has on the efficiency of bacterial transformation.
Agar was prepared for eighty plates with X-Gal which, when metabolized by β-Galactosidase produces a blue dye that coats the E. coli cell. Two-thirds of the plates also had ampicillin added to their media. The pGAL plasmid codes for ampicillin resistance and successful transformation should allow for blue colony growth on plates with both X-Gal and ampicillin in their media. Three groups were divided out of 80 plates: Control (X-Gal/noAMP/no pGAL), Experimental 1 (X-GAL/AMP/no pGAL), and Experimental 2 (X-Gal/AMP/pGAL). Within each group half were radiated under UV-C light for 30 seconds each.
The hypothesis, E. coli samples not exposed to UV light will be have a higher rates of pGAL plasmid transformation success indicated by a higher blue colony count, was supported. E. coli plated with pGAL plasmid not exposed to UV light had higher transformation efficiency than E. coli plated with pGAL plasmid exposed to UV light.