Sea Jellies possess the mystifying ability to glow in the dark. This is due to the presence of the Green Fluorescent Protein. Stanley Cohen and Herbert Boyer were the first scientists to create a recombinant DNA plasmid. They took a gene that coded for a protein in frogs and inserted it into a bacterial plasmid. They did this using a restriction enzyme and a process called heat shock which forces the recombinant DNA through the bacteria's membrane. They then tested the bacteria and it had been tricked into producing frog protein.
We will use these same principals in an attempt to insert the pGLO gene into E. coli bacteria to make them glow in the presence of UV light. To do this we must first create the recombinant DNA using a restriction enzyme and then pipetting the Gene of Interest into the newly cut DNA. We will force them to accept it using the heat shock method of making them very cold then very hot rapidly. Once the new plasmid is in the E. coli we will place it on 4 plates. One will contain only food and will have un-altered bacteria on it which will be the experiment control. Another will have bacteria without the gene and grown on a plate with Ampicillin on it which the bacteria will be vulnerable to without the GOI. The next plate will have the gene with Ampicillin added to it, the bacteria should be resistant to the anti-biotic because of the ampR gene inserted into the plasmid with the GOI. The final plate will have The gene ampicillin and aravnose which is what will cause the bacteria to glow in the presence of UV light.
The bacteria were able to grow and the results were as expected with only one of the plates able to glow, and one plate with no bacteria on it due to its inability to withstand the ampicillin.
We will use these same principals in an attempt to insert the pGLO gene into E. coli bacteria to make them glow in the presence of UV light. To do this we must first create the recombinant DNA using a restriction enzyme and then pipetting the Gene of Interest into the newly cut DNA. We will force them to accept it using the heat shock method of making them very cold then very hot rapidly. Once the new plasmid is in the E. coli we will place it on 4 plates. One will contain only food and will have un-altered bacteria on it which will be the experiment control. Another will have bacteria without the gene and grown on a plate with Ampicillin on it which the bacteria will be vulnerable to without the GOI. The next plate will have the gene with Ampicillin added to it, the bacteria should be resistant to the anti-biotic because of the ampR gene inserted into the plasmid with the GOI. The final plate will have The gene ampicillin and aravnose which is what will cause the bacteria to glow in the presence of UV light.
The bacteria were able to grow and the results were as expected with only one of the plates able to glow, and one plate with no bacteria on it due to its inability to withstand the ampicillin.
