Pglo Essay

The hypothesis above tested the insertion of the pGlo gene to see if the bacteria, E.Coli, will reproduce and grow in the presence of ampicillin and to see if it will cause a green fluorescent glow. (PGLO™ Bacterial Transformation Kit,2017). Based upon the results from this experiment the hypothesis did support the hypothesis and that the presence of the pGlo gene inserted into the E.Coli did cause for growth and fora fluorescent glow to occur. In the experiment, the petri dishes that contained no pGlo (-pGlo) did not show any reproduction nor did a green glow appeared in both dishes. Unlike the two petri dishes, that contained the pGlo gene and ampicillin, the data data showed both reproduction and a glow in the petri dishes.

Coli cells would engage in the pGLO plasmid and fluoresce. one funding did not support the hypothesis. One plate that contained arabinose did not no glow because the cells did not engage in the pGLO plasmid, due to the lack of ampicillin on that plate. The presence of ampicillin is a motivator for the E. Coli cells to engage in pGLO, which also contains beta-lactamase. According to the student manual, “Beta-lactamase acts against ampicillin, protecting the cell from the antibiotic’s harmful effects. Without the threat of ampicillin, the cells have no reason to take up the pGLO plasmid, and therefore the presence of arabinose has no effect on whether the cells fluoresce or not” (Lab

In this experiment the objective was to transform E. coli with the pGLO plasmid and calculate the transformation efficiency. The hypotheses were that the plate with only LB agar and untransformed E. coli would grow a lawn; the control plate of untransformed bacteria with LB and ampicillin would experience no growth; the transformed plate with just LB and ampicillin would grow colonies of bacteria but it would not glow green under UV light; and the transformed plate with LB, ampicillin and arabinose would grow colonies that would glow green under UV light. The results found supported each of these hypotheses as the bacteria grew as predicted. The

One use of GFP is as a component of the plasmid pGLO. pGLO is made through recombinant DNA technology and consists of the GFP gene that codes for green fluorescence, a gene (bla) that codes for the enzyme beta-lactamase which produces bacteria resistance to the beta-lactam family of antibiotics of which ampicillin is one such antibiotic and lastly AraC which binds the arabinose thereby regulating expression of GFP. GFP will not be expressed unless the carbohydrate arabinose is present. Plasmid is small pieces of DNA that is distinct from a cell's chromosomal

The main objective of this experiment was to attempt bacterial transformation, and alter the initial plasmid of DNA to one that expressed GFP, and was therefore able to glow under the UV light. The methodology we used to complete this process was bacterial transformation which is a widely used method where foreign DNA is introduced into a bacterium, which can then amplify, or clone the DNA. Our results were that there was growth on all of the plates except for the-pGLO plate with LB and ampicillin. The results for the fluorescence of the plates was that only the +pGLO plate with LB, ampicillin and the arabinose sugar glowed.

Answer the following questions on another piece of paper in complete sentences using no unidentified pronouns.

Figure 3 shows the growth of three differently treated E. Coli plates. -pGLO plates are both controls. -pGLO with only LB experienced the most grow because

Day one’s lab focused on bacterial transformation. This process is the way in which new DNA is introduced into a bacteria cell. In this process, the bacterial cells need to be made permeable through treatments by way of heat or by mixing with different salts. Once the bacteria cells are made permeable, the plasmid can introduce the genetic instructions used for GFP in the cells. Those that are made permeable have a much higher tendency to express the protein.

DNA encodes the genetic instructions for cells to carry out their daily activities. DNA can come in many forms; plasmids for example are small circular DNA molecules found in most bacterial cells. Though plasmids may not be essential for the life of bacteria, it can give cells resistance in foreign environments. For the purpose of this experiment, an ampicillin-resistant plasmid is introduced to E. coli. This is done through a process of genetic engineering called transformation. Transformation works through the uptake, incorporation, and expression of a foreign gene to alter the genetic code of a cell. Three conditions are needed for successful transformation: a host, a vector, and a technique to identify the transformed cells. E. coli is used in this experiment as the host (E. coli is commonly used in biotechnology due to its rapid rate of growth and short reproduction time). A vector mediates the transfer of foreign DNA into the host cell. Plasmids are commonly used vectors that will also be used in this experiment. The procedure of tagging is used in this experiment to differentiate the transformed cells from those that were not. The learning objectives of this experiment are to: observe the process of bacterial transformation in an experiment; and demonstrate a change in phenotype due to uptake and expression of the genes in a known plasmid.

Plasmids are the extra-chromosomal DNA in which most bacteria contain in addition to their bacterial chromosome. Plasmids can be used as vectors to transfer DNA into a host bacterium. A nonpathogenic strain of Escherichia coli (E. coli) HB101;K-12, is the host bacterium used to take up the plasmid with the imported gene to be observed. pGLO is a recombinant plasmid that consists of a gene that codes for Green Fluorescent Protein (GFP) and also contains a gene, beta-lactam antibiotic (bla), that expresses antibiotic resistance. GFP is a protein that is produced by the jellyfish Aequoria victoria and has the ability to fluoresce green when exposed to ultraviolet (UV) light. The gene for GFP can be turned on in the transformed bacterial cell in the presence of the sugar, arabinose. The bacteria that contain the beta-lactam antibiotic have the ability to create beta-lactamase, which is an enzyme that hydrolyzes the beta-lactam rings of antibiotics which penicillin derivatives (among a broad range of others) contain, deeming them incompetent. Beta-lactam antibiotics inhibit the peptidoglycan layer of the bacterial cell wall and because E.coli is a gram-negative bacteria, its peptidoglycan layer is easily susceptible to inhibition.

In the scientific community, there have been many advancements in the technologies and processes used to help us understand the truth of the world around us. The invention of the electron microscope by Ernst Ruska and Max Knoll in 1933 is just an example of one of these advancements. There is another advancement that has roots over one hundred and sixty million years old, and that is the green fluorescent protein (GFP). The fluorescence from GFP molecules has made it feasible to look through a microscope at a spatial resolution higher than the diffraction limit. The green fluorescent protein was first found in the Aequorea Victoria, or the crystal jellyfish. It is a bioluminescent hydrozoan jellyfish, or hydromedusa (1). In the crystal jellyfish, there are two proteins that are responsible for its bioluminescent, aequorin and GFP. They are both located in the photo-organs of the jellyfish. The jellyfish releases calcium ions which the aequorin binds to. The process of binding gives off a blue light, which is then absorbed by the GFP. The blue light absorbed is then converted into the green light that GFP is named for (2). At the moment there is no apparent reason for the bioluminescence displayed by the Aequorea Victoria since it has not been observed being used as a defense mechanism or as a signalling mechanism. Osamu Shimomura was the first person to isolate the GFP. He started to study the bioluminescence of crystal jellyfish at Princeton University in 1960 and he and

The plasmid was then inserted into a strain of E. coli and grown. After incubation the E. coli strain was found to fluoresce upon illumination with UV light inside of the growing cell, without the addition of A. victoria products (figure 1). Partially purified GFP protein from the bacteria was then obtained and found to posses an identical excitation and emission spectrum to the original A. victoria GFP, as seen in figure 2. This result suggested that GFP did not require additional enzymes/cofactors from A. victoria to fluoresce. Chalfie, using the same technique for GFP insertion in E. coli, inserted the gene into C. elegans under the control of the mec-7 gene, which encodes β-tubulin in C. elegans’ six touch receptors. The GFP again fluoresced when exposed to blue light, but this time the fluorescence was limited to the β-tubulin transcribed by the mec-7 gene, as seen in figure 3. Martin Chalfie, from the results obtained during his experiments was able to conclude that GFP isolated from A. victoria could fluoresce in living cells when exposed to blue light, without cofactors or enzymes form A. victoria. He also showed that gene expression could be traced using GFP, by measuring different fluorescence levels in C. elegans larvae as they

IVF treatment must occur before PGD. Embryo development occurs on day 3 when a small hole is made in the soft shell of the embryo (Zona Pellucida) using a laser. On day 5, one or more cells are removed from each embryo (embryo biopsy). The embryos are then frozen and cells are sent to a PGD laboratory for analysis. The cells are tested from each embryo which informs them which embryos are normal. Normal embryos are thawed and transferred to the woman later.

Genetic transformation is the process involving the insertion of new DNA into a bacteria cell such as E.Coli. While the inserted DNA contains a large chromosome, it also contains small circular pieces of DNA called plasmids. The injected plasmids hold specific genes as they are encoded through genetic engineering. Genetic engineering is described as “the direct manipulation of genes for practical purposes” (Urry et. al., 2016). The DNA injected directs the synthesis of proteins due to genetic expression and explains how bacteria is given the ability to resist antibiotics through the use of the pGLO plasmid. The “pGLO plasmid has been genetically engineered to carry the GFP gene which codes for the green fluorescent protein” (Biorad). This

Hypothesis: If genetic transformation involves the insertion of a gene into an organism in order to change the organism’s trait where genes can be moved from one organism to another with the aid of a plasmid and pGLO plasmid encodes for a gene resistant to the antibiotic ampicillin as well as the gene GFP which allows the bacteria, E. coli, to express the glowing gene by producing the fluorescent protein if switched on after having undergone transformation by adding sugar arabinose – which is a nutrient for the cell -, then when the procedure of the pGLO transformation lab is completed as instructed in the student manual, the agar plate +pGLO Lb/ampicillin will produce colonies of bacteria that do not glow, -pGLO LB/ampicillin will not produce any bacteria, and –pGLO LB (constant) will produce a lawn of bacteria that does not glow (instead appearing as a