Playing God: Transforming Bacteria with Sea Jelly Fluorescence Gene

   Genetic transformation, with diverse applications like bioremediation, gene therapy and genetically modified foods, is one of the most important techniques in the biotech field. In this experiment, we will transform bacteria with a gene from a sea jelly that codes for Green Fluorescent Protein. If everything goes as planned, our bacteria should glow under UV light.
   A bacterium is a prokaryotic single-celled organism with one long, linear chromosome and sometimes a circular plasmid containing specialized DNA. Genetic transformation occurs when a bacterium ingests a foreign DNA strand and expresses it by incorporating the strand into its own DNA. For our lab, we will transform bacteria with a plasmid containing the GFP gene and a gene for ampicillin resistance. The plasmid also contains a gene regulation system, so that the gene for GFP is only expressed in the presence of arabinose.
   Bacteria lack a nucleus and contain all their vital genes in the linear chromosome. Because they multiply rapidly, bacteria colonies can become visible on agar overnight. To make the bacteria competent for transformation, we must alther their cell walls to allow the passage of DNA. We will do this by bathing the cells in calcium chloride and shocking them with heat. The CaCl neutralizes the cell wall and prevents it from repelling the negatively charged DNA. The heat shock therapy stresses the cell and causes it to take in foreign DNA, in this case the GFP plasmid.
   Transformation will not be successful in all of the bacterial cells. Therefore, we will spread the bacteria on an agar plate containing ampicillin, and only the bacteria that took up the plasmid containing GNP and ampicillin resistance will survive; the untransformed bacteria will die. Finally, the moment of truth. When we turn the lights off and apply the UV light, we hope to see a colony of glowing bacteria.
   Last year when I did this lab, we thought we had done everything correctly, but the bacteria did not end up glowing. It was the ultimate disappointment in the coolest lab of the year, so I am seeking redemption.