Green Fluorescent Protein Lab Essay

This pBlu lab had for purpose to present the changes of the strain of E. coli bacteria due to new genetic information being introduced into the cell. In this experiment we are freezing and heat shocking the E. Coli bacteria that is then forced to take the plasmid DNA. The E. coli then transforms the pBLu plasmid, which carries the genes coding for two identifiable phenotypes. After following the Carolina Biological steps our lab worked well and we able to see some colonies of bacteria on the plates. The x-gal plate showed a significant amount of bacteria to confirm that the pBlu plasmid took over the E. coli strain.

When faced with a crisis of identity, we often stick with those who care in the best position to support us. In Neil Smith’s “Green Fluorescent Protein,” When Max was fighting with Ruby-Doo; he wants to stay with his mom for comfort. GFP is a metaphor for Max’s realization that he is queer. Max is ensuring of his feelings towards Ruby-doo. “In Pop’s arms, the white guinea pig turns a brilliant green…… It’s very weird and really scary. [it’s] kind of beautiful in an unexpected way” [50]. He tried to say same sex love can be weird and scary, but it’s love and love it a beautiful thing. Family plays an important role in max’s life. This is what for example Max’s said “I want to say, don’t go. Not that I’m afraid her date

The pGLO plasmid is engineered to express green fluorescent protein (GFP) in the presence of the sugar arabinose as well as the ampicillin resistance gene β-lactamase (bla) (Brown, 2011). Original E. coli HB101 do not have ampicillin resistance or the GFP gene allowing them to glow under UV light. In this experiment, we transformed E. coli HB101 with the pGLO plasmid by heat shock to make the bacterial cells competent, allowing the plasmid to enter the cell (Brown, 2011). The mixture of bacteria with pGLO plasmid were given recovery time after heat shock, then spread on LB/amp and LB/amp/ara agar plates. The bacteria mixture with no plasmid added were spread on LB and LB/amp agar plates and all four plates were incubated at 37°C for

The color of the bacteria was a whitish color and the colony size is similar both before and after the transformation. The best way to do it is to compare the control of the experimental plates. Cells that were typically not treated with the plasmid could not grow on ampicillin, although cells that were treated with the plasmid can grow on the LB/AMP plate. The plasmid would have to confer resistance to ampicillin. Moving on, the GFP gene is what is glowing in the plate because it was activated by the sugar arabinose. The sugar arabinose and the plasmid DNA are also needed to be present because that is what initially turns on the GFP gene which makes the bacteria glow. Organisms can also turn on and off particular genes for camouflage reasons. An organism would benefit from turning on and off certain

Green fluorescent protein (GFP) comes from the jellyfish Aequorea Victoria is rare proteins with high fluoresce and absorbance. The purpose this experiments is to purify and express a His2-tagged recombinant from of GFP (rGFP) from the E. coli strain BL21(DE3)< pRSETA-GFPUV > through a series of experiments by using Ni+2 agarose affinity chromatography technology. The GFPuv gene (UV-optimized GFP) was over expressed in the E. Coil strain BL21 (DE3) (pLysS) as an n-terminal His6/Xpress epitope tagged bind protein. Then using Ni2+ Agarose affinity chromatography to obtain purification of the crude extract. Then observe under the long wavelength UV light, the activity of the rGFP in the column fraction. Bradford assay was performed to obtain the total protein amount. When calculating the

In the pGLO Bacterial Transformation lab, Escherichia coli is transformed with a gene encoding green fluorescent protein by inserting a plasmid containing the GFP gene, beta-lactamase, and arabinose into the bacterium. Successfully transformed bacteria will grow in the presence of ampicillin and glow a bright green color under ultraviolet light. The sugar arabinose is responsible for switching on the GFP gene in the transformed cells, without it, the gene will not be expressed.

to infer that bacterial amylase would break down the starch better and faster than the fungal

Our hypothesis was half right and half wrong in first thinking that the +pGLO plate with amp and ara would have the most colonies of bacteria and glowing bacteria because the most colonies of bacteria were in the +pGLO with only amp. For example, in the experiment, we had 4 plates with E. coli bacteria on them, 2 with the amp, 1 with ara, and 1 just plain bacteria solution. The +pGLO were the only 2 plates that should have been glowing, because they got the pGLO plasmid DNA of a jellyfish and the other two plates didn’t. This proves the hypothesis to true in the glowing part because it shows that we were right in the +pGLO plate having glowing bacteria. Additionally, after our experiment, we show that the +pGLO plate with the amp was the plate with the most colonies of bacteria and the other +pGLO plate with amp and ara only had one colony of bacteria.

Flow Diagram Homework Assignment Name: Preetam Ganti Using the table below, explain the purpose of each of the steps of the Experimental Flow Diagram (Figure 1 from your lab manual – it is provided on the next page) for answering the two research questions guiding this laboratory investigation: 1. Is the bacterial contamination at these three farms due to the same plasmid or different plasmids? 2. What is the frequency of tetracycline resistant bacteria in the beef farm cultures?

Introduction: We are obligated to construct a pGLO plasmid with a gene from a jellyfish that is encoded with Green Fluorescent Protein (GFP). The Green Fluorescent Protein is an antibiotic resistance gene. The bacteria throughout our experiment of the transformation of pGLO is transformed by the existence of ampicillin; which the primary inducer is to express GFP. Using the LB platelets and the LB-Amp-ARA platelets the plasmid

This plate can be used to specifically compare against the plate that contained ampicillin and plasmid DNA. By comparing these two plates, one can see the growth that was able to occur in the plates that contained the plasmid DNA versus the plate that contained no plasmid DNA. In trial A, these two plates cannot be compared because both plates lacked growth (Table 1). In trial B they can be compared because the plate containing the plasmid DNA was able to grow (Table

The technique that allows scientists to follow with their own eyes the dynamic movements of specific proteins as they occur within the living cell is the green fluorescent protein. This is a protein that is extracted from only a few jellyfish. This diffuses a green fluorescent colored light. The green fluorescent protein can be blended to the protein and can work normally and so does the protein itself that it is binded to. The protein is not affected and can be moved and transported throughout the cell and can then be seen.

The purpose of the GFP tag on the gene was to detect the proteins produced by the gene through an anti-GFP antibody. Yeast strains were grown in liquid cultures at either 30°C or 42°C. Proteins were extracted from the yeast strains and amount of total protein in each extract was determined through colorimetric protein assay. An SDS-polyacrylamide gel electrophoresis was conducted. Separated proteins from the gel were transferred to a positively charged nitrocellulose membrane and the blot. Blot was then stained with Ponceau stain to detect total protein. Ponceau stain showed that all lanes contained yeast extract and showed that proteins stayed intact through the experiment. Lastly, primary antibody was added to detect proteins that were bound to the GFP tag in the western blot. After blots were exposed to light, results were obtained on a

As we know in UV spectroscopy the sample is between the source and the detector. So the light starts from the source and goes through the sample and ends in the detector. On the other hand in fluorescence spectroscopy the light is imparted at all angle as a result the detector misses a lot of light. This is a reason that fluorescence spectroscopy is more sensitive that UV spectroscopy.

coli cells can take up foreign DNA plasmid and that will allow the cells to grow on medium containing ampicillin. The plasmid contains an ampicillin resistance gene along with E. coli origin of replication and lacZ sequence containing multiple cloning sites. In the presence of lactose, the lacZ gene produces β-galactoisdase enzyme that metabolizes the lactose. If the foreign DNA inserted within the lacZ gene, then no alpha complementation would occur so functional β-galactoisdase enzyme is not produced. As a result, white colonies of bacteria will form on agar gel. If an insertion occurred outside the lacZ gene, then the cell will be able to produce β-galactoisdase. As a result, blue colonies will form on the plate. The blue-white screening method will allow for the detection of alpha