In this experiment, the idea is to become familiar with the transformation of cells. A well thought out procedure, involving a heat shock procedure, a good antibiotic, an inducer known as arabinose to show the newly expressed DNA by a visible fluorescent glow, and a stable control group is what contributes to this experiments thoroughness. It is predicted that the four agar plates will all yield different forms of growth, with different coloration and colony number. It is also predicted that the agar plate media containing the arabinose will grow a green fluorescent color and the (-)pGlo plate with ampicillin will exhibit no growth at all, due to the lack of the plasmid that is resistant to ampicillin. The prediction for the other two plates lacking arabinose is a growth of colorless colonies. Multiple medical experiments are credited towards transformation, especially ones involving cancer and the transformation of normal cells to neoplastic and cancerous cells.
The general purpose of the Transformation Lab was to observe the difference in bacterial growth under differing media conditions to help understand the process of transformation and how it contributes to the life of living organisms. "Transformation" is what occurs when a cell receives and expresses a new piece of DNA that was otherwise foreign to it before . This type of experiment demonstrates what happens when a cell transforms and expresses a gene it once did not have, which is basically the gene that makes it glow fluorescent green. This particular glowing gene comes from the jellyfish Aequorea victoria. This glowing gene is taken from the jellyfish and the E. coli takes in the GFP protein from the jellyfish DNA, transforms to accept the glowing gene, and will then provide its own fluorescent green glow under UV light when it begins to express the gene . People may wonder why this type of study is important, besides making things glow in the lab. Transformation is a big part of medical studies, especially with the transformation of normal cells into malignant ones. Cancer is a really big part of why microbiologists study the transformation of cells. It is also a big part of helping sick people by helping them transform their diseased cells into healthy ones donated from a healthy person. This is a new idea that is being explored known as "gene therapy" . Aside from the important things, the idea behind this particular transformation lab is to create a visual on how transformation is done. The green fluorescent glow provides the perfect visual for a transformed gene. It also provides a visual for the way some bacteria become resistant to certain antibiotics. The plasmid taken in by the E. coli bacteria also codes for a resistant to the antibiotic used, which is ampicillin. Through the control group, it is seen that ampicillin kills any E. coli bacteria that has not mutated for resistance to the antibiotic. But, when the bacteria transforms to express the plasmid, which does have a resistance to ampicillin, bacteria grow as normal. This demonstrates how some bacteria get around antibiotics and develop a resistance; they transform and express a new gene that is not affected by that particular antibiotic. Transformation experiments help humans in so many ways and to be able to re-create this cell-to-cell miracle is a fantastic jumpstart for all the things we still do not understand about the microbiological world.
Materials & Methods
Materials needed for this experiment included an E. coli starter plate, 4 agar plates, transformation solution of calcium chloride, LB nutrient broth, Inoculating loops, pipets, foam microtube holder float for (+) and (-) pGLO microtubes, container of crushed ice, rehydrated pGLO plasmid, a 42 degree celsius water bath and a thermometer, and a 37 degree celsius incubation space .
After receiving the two test tubes, label one (+) pGLO and the other...