1. Transformation involves the transfer of genetic information into a cell by directly taking up foreign DNA from its surroundings. This DNA is then incorporated into the host cell’s own DNA. This transformation usually occurs within plasmids, small circular DNA molecules separate from its chromosome. After the recipient cell is infected with the DNA, the cell will move on with replication, producing offspring with traits encoded by the plasmid. These plasmids may replicate with the chromosome, or independently. This is how diseases are commonly spread, since one little bit of DNA can affect the entire organism thanks to duplication.
2. E. Coli are ideal organisms for molecular geneticists to manipulate because it can easily be grown in suspension culture in mediums such as Luria broth or on agar. Also, E. Coli has a relatively small genome, containing only about five million DNA base pairs. By chemically and thermally treating E. coli cells, they can artificially be transformed. Naturally, these cells do not possess the natural system needed for transformation. After treatment, they become receptive to an insertion of foreign DNA contained in a plasmid.
3. After treating the E. coli cells, plasmid containing firefly genes can be inserted into the cell. What this will do is create recombinants in which properties of the original cell and the firefly cell will be exhibited. In order to insert the plasmid into the cells, we will use a sterile inoculating loop to remove a loopful of the E. Coli from the surface of the agar. We will then place the loop into calcium chloride and twirl it rapidly, releasing the bacteria. Then, using a pipette, we will add 10µl of the pBestLuc solution to the mixture.
4. Luciferase is the enzyme responsible for the luminescent glow of the firefly. It causes the glow by catalyzing a reaction between the chemicals luciferin and ATP in the presence of oxygen and magnesium. After injecting the cells with this enzyme, and allowing them to sit for 24 hours, we will be able to observe whether or not transformation was successful based on whether or not our bacteria will glow. 5. The way in which we will calculate the efficiency of the transformation reaction is by comparing our results with those of our classmates. If most of the groups did not have successful transformation, it is safe to conclude that the efficiency is relatively low. On the otherhand, if all the groups see that the bacteria has taken on the new trait, than the efficiency is relatively high.
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Topic: Glowing Bacteria Lab
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