Genetic Transformation Lab Report

The purpose of the PGLO lab was to be able to perform a procedure known as a genetic transformation. We used a procedure to transform bacteria with the gene that codes for a Green Fluorescent Protein (GFP). The actual source of the GFP gene that we used in this complicated experiment is the bioluminescent jellyfish Aequorea victoria. This protein causes the jellyfish to glow under a UV light that was provided in the dark. After the transformation procedure, the bacteria showed their newly acquired gene from a jellyfish and produced the fluorescent protein, which as a result, causes it to glow. If the bacteria glowed in the dark, that was the initial sign that the experiment was successful.

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.

Transformation is the transfers of virulence from one cell to another, through the transferring of genetic material. It was originally postulated in 1928 through the works of Federick Griffith, a British microbiologist. Griffith observed that the mutant form, non-virulent form, of the bacteria Streptococcus Pnumoniae could be transformed into the normal, virulent form, when injected into mice along with heat killed normal forms. He concluded that somehow the information the dead virulent form had transformed the mutant form into a virulent form.

Scientists can study and manipulate genes by using precisely engineered plasmids, According to ADDGENE, a nonprofit plasmid repository, plasmids “have become possibly the most ubiquitous tools in the molecular biologist’s toolbox” (“What Is a Plasmid?”). In this experiment, we will use pGLO to genetically transform the bacteria E. coli. pGLO is a genetically engineered plasmid that carries the reporter genes for both green fluorescent proteins (GFP) and the ampicillin resistance. GFP, a protein typically found in the bioluminescent jellyfish Aequorea victoria, exhibits a bright green fluorescence in the presence of blue to UV wavelengths (Mecham); the protein absorbs UV light from the sun and emits it as a lower energy green light, exemplifying the second law of

Genetic engineering is used in health treatments, agricultural applications, and environmental solutions. Genetic transformations incorporate foreign genetic material into the DNA of a different organism via a vector, which carries the genetic material. Plasmid DNA is small, round, and autonomous, due to its origin of replication. In biotechnology, plasmids carry beneficial genes, such as antibiotic resistance, and also a reporter protein, in this case, Green Fluorescent Protein

Genetic engineering and transformation is the “modification of an organism's original genetic composition by artificial means”(Genetic Engineering 2016). Genetic engineering and transformation involves the swapping of traits and genes between organisms to create a genetically modified organism or GMO. Genetic engineering has it’s advantages which include the prevention of disease by isolation of the gene that causes the disease, being able to take the “desirable traits” of one organism and implanting them into another, and genetic engineering can increase diversity in plants and animals by creating alleles that can be given to different species of organisms according to the Asia-Pacific Economics Blog (Apecsec.org 2014). Unfortunately, genetic

Genetic Transformation in its simplest definition is the alteration of a cell when foreign material such as DNA is introduced through its cell wall, altering its genetic makeup. Genetic Transformation is significant in modern society because of its effect on the agriculture industry and crop production throughout the world. Scientists and farmers everywhere are constantly looking for ways to increase yield and quality of produce, which they have found possible with Genetic Transformation. The agriculture industry is just one common example of an industry that utilizes Genetic Transformation in order to achieve their goals and improve productivity. This process occurs through multiple techniques known as projectile bombardment,

The first genetic engineering experiment was conducted in 1973 by Herbert Boyer, of the University of California at San Francisco, and Stanley Cohen, at Stanford University. The duo had successfully combined and replicated genetic information from entirely different species also known as recumbent DNA (“Genetics”). Genetics has made remarkable progress throughout decades of innovation and discovery and will continue to be a bewilderment that is to be solved. The first decade of the 21st century has been one of the most inventive in this field

If genetic transformation allows the insertion of a gene into an organism to gain the trait that the inserted gene codes for, then when 250 µl of transformation solution(CaCl2) and 2-4 colonies of bacteria, E. coli, are added to two micro test tubes, labelled +pGLO and –pGLO, with a loopful of pGLO DNA solution in the +pGLO tube, and are incubated for 10 minutes, then transferred to a 42ºC heat bath for exactly 50 seconds, and then incubated in ice again for 2 minutes for a heat shock, and then have 250 µl of LB nutrient broth, and lastly 100 µl of the solutions spread across the surfaces of 4 LB nutrient agar plates, LB/amp, LB/amp/ara, LB/amp, and LB, with +pGLO, +pGLO, –pGLO, and –pGLO used, respectively, the plates with the –pGLO solution will not glow under a UV light after they have

The process of genetic engineering has evolved throughout the past several decades. In the process, the organism’s DNA is modified by combining foreign genetic material into the original DNA strand ("UNL's AgBiosafety for Educators."). In order to insert the

Transformation is the process of allowing a cell to be able to accept a gene into itself and allow the gene to become part of the cell 's whole, thus changing its form and expressing the gene 's function. This process was first discovered by the British bacteriologist Frederick Griffith, who was able to transform Streptococcus pneumoniae into a different strain of Strep. in 1928 (1928). This discovery was pivotal in the way we understand gene expression and its engineering. An example of genetic engineering using transformation that applies to everyday life is the synthesizing of human insulin, which is a peptide hormone that helps to regulate the amount of sugar that runs through the bloodstream. In the early 1900s, insulin from other animals was used to treat diabetes, but human insulin is now able to be easily produced in bulk through genetic engineering, and has largely replaced animal insulin for the treatment of diabetes. Insulin is made from cells in the pancreas, and the extraction of insulin from animals used to require taking it directly from the animal 's pancreas glands. It was not until 1977 that synthetic human insulin was genetically engineered by Arthur Riggs and Keiichi Itakura, and was achieved by the process of transformation using Escherichia coli as a host (1977), which allowed for the easy reproduction of human insulin. Using this method, the production of human insulin became large-scale, faster, cheaper, and more accessible.

Bacterial transformation is the process of moving genes from a living thing to another with the help of a plasmid.The plasmid is able to help replicate the chromosomes by themselves; laboratories use these to aid in gene multiplication. Bacterial transformation is relevant in everyday lives due to the fact that almost all plasmids carry a bacterial origin of replication and an antibiotic resistance gene(“Addgene: Protocol - How to Do a Bacterial

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.

Genetic transformation is when an organism is given genes that it is able to express from the foreign DNA (Weedman 111). Genetic transformation is used widely today, especially in agriculture where organisms are genetically altered to meet the food demands from corns color to squash size. When transferring genes you need a competent cell which is the cell that is going to express the foreign DNA (Weedman 111). There are three ways to transfer DNA, transformation where the competent cell takes DNA from the surrounding environment, conjugation where two bacteria join and transfer DNA, and then there is transduction where virus or vector carries DNA from a foreign organism to another (hammiverse.com...na). In our experiment we used transduction, using a vector. There are three ways to make the competent cell permeable. There is projectile bombardment when a gene gun is used to coat the competent cell with DNA, there is electroporation when electric pulses are used to increase permeability and then there is heat shock when you rapidly change the temperature of the environment the competent cell is in from low to high temperatures making the permeability of the cell membrane more profound (Weedman 111). We used the heat shock method in our experiment to increase the permeability of E.coli. In the case of our experiment the competent cell is E.coli. E.coli is known for its ability of horizontal gene transfer which is the transfer of genes from a foreign organism and is not

GFP is a widely used tool in the field of Molecular biology and Cell biology. It involves emission of fluorescence under Ultra Violet light which allows for direct investigation into the inner working of cells. Green Fluorescent Protein was first isolated from the jelly fish Aequorea victoria by Osamu Shimomura (Shimomura et al., 1962). Since its discovery, it has become useful in the field of science. The GFP chromophore is formed from tri peptide in the primary structure of GFP. Its fluorescence is turned on when exposed to molecular oxygen. The gene of GFP has been introduced into many bacteria, yeast, fungi, plants and humans (Amsterdam, A. et al., 1996) and it is still gaining rapid ground in the field of biological science.