Bacteria Pathogens

This is proven by the survival of the bacteria in the LB/amp +pGLO dish, as the same bacteria that lacked the modification were killed by the ampicillin. However, we were unsuccessful in genetically transforming bacteria to be fluorescent. This is because in the LB/amp/arbo +pGLO dish, we were unable to locate any bacteria.

The pGLO plasmid will transform the E. coli bacteria with a gene called GFP that codes for the Green Fluorescent Protein in the genetic code. GFP was discovered in the jellyfish, Aequorea victoria as a green fluorescent light emitted from the jellyfish. It was typically seen in the dark upon its activation and since then has been used in studies relating to genetic transformation. (Chalfie and Tu 1994) The majority of the studies test the many different factors that are required in the transformation of pGLO which will determine the functionality of GFP in the E.coli bacterium. The first experiment in transforming GFP and E. coli was completed in 1994 by Chalfie and was further refined the same year. The experiment proved the importance of using restriction enzymes, and DNA ligase in the process of transforming GFP to identify arabinose as the primary activator, and to identify the ampicillin

Genetic transformation occurs when genes are inserted into another gene to change the organism’s trait (Weedman2016). In this experiment, we proceeded to transform the E. coli bacteria with a gene that contained green fluorescent protein. The green fluorescent protein is used in experiments because it beams a green color under a UV light (Chalfie2008). Typically, it is used to mark the expression of genes, which is why it serves as the symbol for all gene expressions (Tsien1998). In the experiment, we will be using pGLO as the organisms that will transmit the disease, otherwise known as a vector. The pGLO in the experiment

To start the experiment two small plastic tubes were labeled negative pGLO and positive pGLO as the first step in this experiment. A pipet was then used to move 250 microliters of calcium chloride into each tube and the tubes were iced. Bacteria was added using a new sterilized loop each time to the positive and negative pGLO tubes. The loop was twisted around in the tube to ensure that the bacteria was evenly mixed into the solutions and the tubes were iced once again. Another loop was dipped into the tube containing the plasmid and it was removed when there was a visible slight film of residue. The plasmid was dipped into the tube that was labeled positive for the pGLO gene. The two tubes labeled positive pGLO and negative pGLO were iced

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

The objective of this experiment was to genetically transform E.coli cells to express ampicillin resistance and to produce the green fluorescent protein by using the pGLO plasmid. It was hypothesized that only the cells with pGLO DNA added to the solution would be able to survive in the same environment as the ampicillin, and that only the cells grown in the plate with arabinose would fluoresce bright green. The results supported the hypothesis, showing that the +pGLO LB/amp/ara plate was the only plate that had fluorescent cells and grew. Additionally, that +pGLO cells were the only ones that grew on an LB/amp plate.

Coli bacteria, and mix it with the plasmid. We can select the bacteria that are resistant by introducing ampicillin. Bacteria that are not resistant to ampicillin dies. The screening of the bacteria with the resistance occurs when the bla gene turns on to produce beta lactamase which will kill the antibiotic ampicillin. After this, we can get the plasmid into the bacterial cell through the process of horizontal gene transfer through transformation. This can be done by a cold treatment in an ionic solution, then a heat shock where it increases membrane fluidity to take up the plasmid into the bacteria cell. In order to know if the EPAS1-TD gene product was produced by the bacteria, we have to perform a Western Blot. We do this by extracting the protein and run it through gel electrophoresis to separate the proteins by their size. We then label the proteins with the colored probe based on their size. We can detect the EPAS1-TD gene being produced based on the colored

Title Genetically transferring the GFP (green fluorescent protein) gene into E. Coli via heat shock Introduction Genetic transformation is the process by which desirable genetic material is extracted from one organism and forcibly placed in a different organism’s genetic complex. By adding foreign genes of organisms into different organisms, the genetic diversity of organisms is increasing exponentially and genetic transformation is one, if not the best, way to produce new alterations of genes as well as improve the functionality of other, already existing, genes. Although there are many different ways by which genetic transformation can be done, heat shock transformation was the method used in this lab. Heat shock is used to increase the

The purpose of this lab is to observe bacterial growth under various conditions including the transformation of bacteria; to understand how the process of transformation occurs.

Geneticists continue to look for ways to genetically alter genes and transformation to better the lives of other people. In this experiment the goal was to understand the transformation process and how genetically engineering bacteria could possibly be useful. The transformation process involved 5 steps: pre-incubation, incubation, heat chock, recovery and growth, and selection. Four different plates were used to analyze the effects of the transformation. The plates were as follows: plate 1 contained lysogenic broth (LB) and ampicillin (amp) with plasmids (pGLO); plate 2 contained LB/amp/arabinose sugar with pGLO; plate 3 contained LB/amp without pGLO; and plate 4 contained LB without pGLO. The results obtained proved our hypotheses to be correct.

Genetic transformation is the change caused by genes. This transformation includes the insertion of a gene into an organism, changing one of the organism’s traits. There are many other uses for genetic transformation including the altering of plant genes coding for frostbite, pests and spoilage resistance. It can also be used to digest oil spills and even alter in gene therapy to transform sick cells into healthy ones. This particular experiment will include the transformation of the bacteria with the GFP ( Green

Bacterial transformation is one of the main ways scientists are genetically engineering cells. The reason why bacteria is getting used for these experiments is because bacteria’s genes are very versatile. For the purpose of this experiment we used the bacteria Escherichia Coli, the reason for this is that this specific bacterium is resistant to antibiotics. Plasmids were used in this experiment for the purpose of identifying what type of growth will occur also because plasmids are needed in order for transformation to be possible. Plasmid is not the only thing needed, also needed are vectors and a host. The bacteria used, E-Coli, served as the host in this experiment and the plasmids served as the vectors. Vectors are used to deliver foreign

Transformation in bacteria is something that could be essential for survival in a bacteria. In order to perform this transformation naturally a bacterium must considered competent, otherwise it must undergo an artificial transformation. Being a competent cell means that the bacteria can take up DNA from its environment naturally (5). Those that are not competent such as Escherichia coli that are not naturally competent can be tested with an artificial transformation, such as what we will use in this experiment. Methods used can obtain things such as chemical mutagens or radiation (1). The gene used for the transfer is the GFP gene (Green Fluorescent Protein), which gives an illuminating appearance under a UV light when conducted properly,

The heat shock treatment increases the rate of successful transformation by making the bacterial plasma membrane more permeable to the plasmid solution. During the experiment performed in the Life 102 Lab we took a sample of Escherichia Coli (E. Coli) bacteria and transformed it using a gene that codes for green fluorescent protein (GFP). (Weedman 2016). The GFP originates from a bioluminescent jellyfish, and the protein is what allows them to glow. The GFP gene is only visible in the presence of arabinose.

Jellyfish (Aequorea victoria) naturally produce GFP (Zupen et al. 2004) allowing for extraction of the organisms genes and protein to engineer a bacteria that can produce GFP. Scientists are able to blind a bacterial species for example E. coli with GFP by cloning the genes found within jellyfish that produce Green Florescent Protein and insert it into a chosen bacterial species (Zupen et al. 2004). Through the expression of the GFP gene in other organisms, synthetic biology can be