E. Coli Bacteria Lab Report
In this lab, genes for a fluorescent green protein (GFP) and antibacterial resistance (ARG) were inserted into E. coli bacteria. E. coli bacteria was resuspended in an ice-cold CaCl2 solution. DNA containing GFP and ARG was added to half of the cells before they were “heat shocked” in an ice bath and hot water. The heat shocking made the bacteria’s cell membrane more porous, so the DNA could enter. Recovery broth was added to the cell suspension, and the bacteria was placed in warm water for about thirty minutes (see Results and Discussion, paragraph 2). This recovery period let the bacteria repair their cell membranes and express the added genes. Lastly, the transformed E. coli were placed on agar plates and allowed to grow overnight. One agar plate only contained nutrients (-DNA), two contained nutrients and ampicillin, (-DNA/Amp and +DNA/Amp), and one contained nutrients, ampicillin, and IPTG, a protein that caused the GFP to express a glow.
After completing the lab, it was discovered that the ARG gene creates a resistance to antibiotics, like ampicillin, and that bacteria can take in new genes. The control plate, or -DNA, grew over three hundred bacteria colonies as there was nothing to stop its
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This was done by adding genes for resistance to antibiotics and a green fluorescent protein to E. coli bacteria. Over three hundred bacteria colonies grew in the -DNA plate, none grew in the -DNA/Amp plate or the +DNA/Amp plate, and one glowing colony grew in the +DNA/Amp/IPTG plate. It was found that organisms can take in and express new genes, as shown by the growth of bacteria despite the presence of ampicillin in the last plate, as well as its glow. Genetic engineering techniques like this one are used everywhere in the real world, for medicine, agriculture, research, and more; glowing bacteria is just the
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