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The purpose of this lab experiment was to see if a person could use a plasmid (R plasmid) vector containing certain fragments of foreign DNA for ampicillin resistance to be used to transform E. coli (Escherichia coli) cells to give the E. coli cells ampicillin resistance. Ampicillin resistance is needed in order for the E. coli to have the ability to survive in an ampicillin environment. Plasmid is a small ring of DNA that carries accessory genes separate from those of bacterial chromosome. Ampicillin is an antibiotic that is derived from penicillin that prevents bacterial growth by interfering with cell wall synthesis. E. coli is a commensal bacterium inhabiting the human colon that is widely used in biology, both as a simple model of cell biochemical function and as a host for molecular cloning experiments. In nature genes can be transferred between bacteria in different ways like transformation, conjugation, and transduction. One-way is bacterial transformation, it is when transfer of genetic information into a cell by the direct uptake of the DNA. Then the DNA is used to transform the cells from the original DNA to take a certain trait. How ever these bacteria can take up DNA only during the period at the end of logarithmic growth. Conjugation is performed between to bacterial cells of different mating types and genetic information is exchanged between the two through pili. Transduction is conducted between two bacteria cells, but this process requires the presence of a virus that acts as a vector in the process.
One could hypothesize that plasmid would be able to successfully be incorporated in the E. coli cell. One would be able to see this by both agar plates without ampicillin and also the agar plate with ampicillin and plasmid would have bacteria growth. However the agar plate with ampicillin, but no plasmid will have no bacteria growth. One could hypothesize this because if the plasmid were incorporated into the E. coli cell correctly the E. coli would have ampicillin resistance.
2 Luria agar plates with Ampicillin(control)
1) First get two microcnetrifuge tubes and mark one of the tubes + and then the other tube -. The tubes with the + is the one that the plasmid will be added to it. Then add 250ml of cold calcium
Terminology mentioned in this term paper
E. coli, Calcium Chloride, DNA,
Drug mentioned in this paper
Keywords talked about in this paper
plasmid, ampicillin, agar plates, bacteria, pipet, calcium chloride, r plasmids, luria broth, the tubes, cell, tube, bacterial growth, Escherichia coli, antibiotic, inoculating loop, solution, antibiotic resistance, the cells, logarithmic growth, one plate, experiment, bacterial chromosome, bacterial transformation, cell wall, penicillin, bunsen burner, Loop 1, cell division, commensal bacterium, disposable, flame, room temp, molecular cloning, hypothesize, minutes, hot water, procedure, broad spectrum, the next one, real life, right away, be incorporated, to survive, results, epidemic, large, mutants, hospital, information, Hotplate,