Biology Paper

443 WordsAug 25, 20142 Pages
Ryan Browne 1. The Endonucleases I used were BamH I and Hind III. From looking at the map of my plasmid I found that the best endonucleases to use were indeed BamH I and Hind III. The gene of interest fell in between both of these endonucleases cut sites on the DNA. On the plasmid, BamH I fell in the middle of the Kanamycin Resistance and Hind III is in between the resistance site and the origin if replication. This allowed for one site of resistance to be fully functional and for the plasmid to still have its sight of origin. 2. Of all the Endonucleases, I did not use EcoR I, Ava II, Hpa II, and Sac I. I could only use two pairs of endonucleases to obtain the gene of interest, first BamH I and Hind III, or EcoR I and Hind III. This immediately eliminates Ava II, Hpa II, and Sac I. After reading the requirements in step D: 1, I found that if I used Eco I, I would either cut out the origin of replication or I would cut both the restriction sites into nonfunctional sites. This only left me with the choice of BamH I, and Hind III. 3. You could put ampicillin in with the E. coli because we would be able to tell the plasmid still had the ampicillin resistance site in it so the bacteria with the plasmid would survive and the bacteria without the plasmid would die. 4. The mRNA was able to pair up with the tRNA creating a sequence out of amino acids. The amino acid sequence was: MET LEU TYR CYS ARG GLU ALA VAL THR GLY GLY LYS PHE THR LEU STOP 5. The polypeptide produced would have been an Adenine when translated. This would have changed the amino acid to a LYS instead of a MET. Therefore the protein would not have been created. 6. The mRNA had one extra base at the end of where we translated it. Therefore the polypeptide chain wouldn’t have a stop codon, this would then create an error with the amino acids and the protein would then not work.. The

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