Introduction: Transforming a gene or genetic information from one organism into another with the hopes that if done successfully the organism with the new DNA will be given new traits is a method known as genetic transformation (Rafter). Genetic transformation is used quite frequently in today’s world, form medicine to agriculture. In this lab we will be inserting a gene into an Escherichia coli bacteria with the help of a plasmid. Escherichia coli bacteria also known as E. coli, is a bacterium that is rod shaped and contains flagella to help it move. The bacterial DNA is circular inside of an E. coli bacterium. E coli. is most known for being found in the intestine of humans and animals but it can also be found in other places such as food …show more content…
coli bacteria new traits. The pGLO plasmid that is being transformed into these cells contains genes that can give colonies of bacteria the ability of antibiotic resistance and a green fluorescent glow. Four different models were prepared and plated on multiple agar plate. After the bacteria was grown for three days in an incubator at 37°C; observations were made and recorded (Table 1). All of the plates were looked at for the amount of colonies grown, if growth was present, and if the colonies gained the ability to glow green. Transformation was successful in the plates where the bacteria consumed the pGLO plasmid. In the first plate that the bacterium was plated on it included the LB broth and of ampicillin antibiotic (amp), 2 colonies were present. The second plate of bacteria was grown with the presence of LB broth, ampicillin, arabinose sugar (ara), and 22 colonies were observed. But a green fluorescent glow of the colonies was only present in plate 2. Plates 3 and 4 were the control plates. The bacteria were not transformed and no growth was present in either …show more content…
Our hypothesis was that if the plate contains only the LB broth the E. coli bacteria would have no antibiotic resistance and would not glow. If the plate contains just LB broth and ampicillin then the E. coli bacteria will have antibiotic resistance only if the plasmid is present. If the plate contains LB broth, ampicillin and arabinose then the E. coli bacteria will glow fluorescent under a UV light and it will have antibiotic resistance. Similar to our expectations our results suggested that our hypothesis was correct. This is due to the fact that n order for the E coli. bacteria to gain either of the traits the plasmid had to be present in the first place, because the GFP gene is inside the plasmid. If arabinose is not in the media in which the bacteria was growing on then the GFP gene could not turn on, thus the bacteria can not glow. This is why the LB/amp/ara plate was the only one to express both traits(antibiotic resistance and glowing). The second part to our hypothesis was that using HIC,GFP would be purified and the final tube would express GFP under an ultra violet light. Contrary to what we expected the final tube nor any of the columns expressed the ability to glow under ultra violet light. This unexpected result may have been
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Exercise 14: Unknown Identification Lab Report The purpose of the study was to identify the unknown bacterium using various biochemical tests in addition to using scientific methods in determining the outcome of the hypothesis. Each biochemical test will help determine the bacteria based on specific characteristics of each organism. I was giving unknown number 232. The first procedure that needed to be done after obtaining unknown bacterial mixture was to isolate the two bacteria in a pure culture using the streak plate method described in Microbiology Laboratory Manual Eight Edition. The material used was trypticase soy agar (TSA) plate, nutrient plate, starch agar, hydrogen peroxide, iodine reagent and microscope.
This experiment was tested on two conditions: +pGLO and -pGLO. +pGLO has the plasmid and -pGLO does not have the plasmid. Along with those conditions, ampicillin is added in the agar plate to destroy the non-transformed cells because scientist want to select only the transformed bacterial cells and the experiment is efficient at ten percent. To resist the antibiotic, the bacteria needs Beta lactamase that provide the enzyme. This allows the bacterial to break down the antibiotic and grow and with the plasmid.
To begin, during this lab experiment, genetic transformation was successfully carried out. After observing the agar plates, it was found that only the plate with ampicillin and no pGLO plasmid did not grow any of the E.coli bacteria. All three of the other plates grew the E.coli bacteria, however it grew differently in each plate. In the control plate where the pGLO plasmid, ampicillin, and arabinose were not present, the bacteria grew in the pattern that it was spread in originally. In the two other plates, bacteria grew in colonies that eventually joined together due to prolonged time in the incubator.
The tube was placed back in incubation for 96 more hours to observe any more positives. 2.10 Catalase Test A trypticase soy agar plate was used and after incubation, four drops of 3% Hydrogen Peroxide was added to the plate to flow over the bacterial growth. A presence of bubbling was observed. 2.11 Starch Hydrolysis
On the first negative control plate, there should have been no growth on the plate with agar and ampicillin and regular E. coli cells. On the negative control plate with just agar and E. coli cells there should have been growth but the colonies would not
But, since no arabinose was present, the bacteria was unable to glow. In the second dish, +pGLO LB/amp/ara, the bacteria was able to glow and grow since there was the pGLO plasmid which contains the gene that is ampicillin resistant, and the sugar arabinose which activates the araC gene which allows the bacteria to glow. In the third dish, -pGLO LB/amp, the bacteria did not glow or
Q1A: What is the mechanism of action of colistin? Colistin is an antibiotic that works best against Gram-negative bacteria. It works by binding to LPSs (lipopolysaccrides) and phospholipids in the outer cell membrane of the bacteria. This, in turn, disrupts the outer cell membrane by displacing cations and leaking the intracellular contents, combining it with outer cellular contents, causing the bacteria to be unable to differentiate the bacteria’s intra and outer cellular contents from one another.
After experiment on microscope under oil immersion, I learned that my Unknown is gram positive. Under the lens, the bacteria appears in purple color. Its morphology is cocci arranged in cluster. However, during decolorizing process, I put too much alcohol to the crystal violet-iodine complex making the color overly removed. That led to the result of my gram positive has slightly redish
The putrid smell of Escherichia coli is one that is immediately identifiable to the few lucky individuals who recognize its scent. It is also an aroma with which I became intimately sensitive to as I shuttled petri dishes of the bacterium in and out of an incubator. While my classmates shied away from the task of handling the pungent bacteria used in our recombinant DNA experiments, I took to the task eagerly, anything that would take me one step closer to my goal of researching. I had the opportunity to learn about lab techniques and cutting edge biology concepts the summer before my junior year, in an extracurricular biotechnology class at Northwestern University’s Center for Talent Development. The class, a three week crash course in the
Contrary to popular belief, Charles Darwin was actually not the first biologist to suggest that species rehabilitated over time into new more developed and advanced species. Darwin alongside Jean-Baptiste Lamarck and a few other naturalists eventually coined the concept of evolution by the end of the 1700s. For most of his life, Lamarck focused on his two proposed mechanisms, of which both were very successful in the science world. As species adapted to the environment and their surroundings, the nature of their species also began to progressively adapt and become more complex springing from simple to more complex beings. Lamarck also hypothesized the idea of spontaneous generations in which he theorized that new primeval life species essentially