Plasmid

The focus of this lab was to identify which plasmid (pFG or pGLO) transformed into E. coli culture. E. coli have the capability to take up foreign DNA from their environment in times of stress. In order for the E. coli to be transformed, the E. coli must first be made competent. Once the culture has been made competent, the cells can transform. The plasmid that was inserted into the E. coli culture contained antibiotic-resistant gene and a reporter gene. A reporter gene is a gene that is easy to…

conducted by using a plasmid containing various genes called pGLO along with the bacterium Escherichia coli. In order for transformation to occur in Escherichia coli, repeated washes in the presence of CaCl₂ followed by heat shock treatment (42°C for 50 seconds) occurred to induce the competence of the bacterium. Successful transformation results were expressed by phenotypic characteristics such as growth in the medium and glowing from exposure of UV light from the pGLO plasmid genes…

whether or not the bacteria would glow under UV light. Hypothesis: If the bacteria with the pGLO plasmid was grown on a plate containing LB and ampicillin then the bacteria will grow but not glow under UV light. If the bacteria with the pGLO plasmid was grown on a plate containing LB, ampicillin, and arabinose then it will be able to grow and glow under UV light. If the bacteria without pGLO plasmid was grown on a plate containing LB and ampicillin then it will not be able to grow or glow under…

10/25/17 Bacterial Transformation Utilizing E. Coli Background Through this lab there were many terms used that may confuse you so before I begin explaining the lab, here’s some background information on the subject of biology. A plasmid is bacteria naturally containing one or more tiny circular pieces of DNA. The two genes being used in this experiment are Ampicillin and fluorescent genes. When reading about the lab, the most common thing used in the report is the “+”…

Transforming Genetic Material To E. Coli Using Plasmids Proteins are created within the body through a process known as central dogma. This process is the transcription of DNA to RNA and the translation of RNA to proteins. The type of protein created relies on what type of messenger RNA or mRNA that was produced during the transcription of DNA. In our experiment, we looked at the specific LacZ gene that codes for beta-galactosidase, which is a protein/enzyme that can break down lactose into its…

in the form of a plasmid called pGLO, was added to a sample of E. coli and a control was created of E. coli without it. The E. coli was then placed in either ampicillin or not as well as with a ribosome known as arabinose. The E. coli without pGLO died in the ampicillin and the ones with pGLO survived after a period of incubation, with the one that had arabinose glowing under ultraviolet light. This was the expected result due to GFP adding a resistance to ampicillin in the plasmid.…

transfers genetic material to the recipient bacterium. During the process of conjugation, one of the bacterium aids as the donor of the genetic material. The other serves as the recipient. The donor bacterium carries a DNA sequence. One strand of the plasmid is transported to the other and the other remains in the original cell. Since…

The ways in which bacteria can become resistant to antibiotics and how this contributes to the global healthcare concern of Antimicrobial Resistance (AMR). Antimicrobial resistance is a prevailing issue since the discovery of the first antibiotic Penicillin in 1928. There are 5 mechanisms which allow resistance to impede new antibiotic development for the last 29 years (WHO, 2016) consisting of mutations in target genes, enzymatic resistance, latency, antibiotic efflux and non-specific mutations…

extracellular plasmid of DNA. It involves introducing the E. coli and allowing it to adapt to the DNA of a new environment. The E. coli must be placed in a tube, into an ice bath, causing the cell to shrink. From there it is transferred to a preheated bath, where the cell is heat shocked, allowing the plasmids ' to enter the cell at a rapid pace, once the cell expands. After the tubes are incubated, they were transferred to six agar plates. There were two types of agar plates, one included the…

Methods and materials Cell culture medium Making cell culture medium, requires two processes, this includes the medium that has PSG and other without the PSG. 15 ml of 10% FBS was added into a culture flask with 1.5 ml 100x hippos. The culture flask was filled with DMEM up to 150 ml and this will be the medium without the PSG and this process completed the medium without the PSG. The same procedure was followed for the PSG medium, 1.5 ml of PSG was added and that completed the medium with PSG.…