Dna Coding For Ampicillin Resistance And Green Fluorescent Protein

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 plasmid pGLO contains an antibiotic-resistance gene, ampR, and the GFP gene is regulated by the control region of the ara operon. Ampicillin is an antibiotic that kills E. coli, so if E. coli, so if E. coli cells contain the ampicillin-resistance gene, the cells can survive exposure to ampicillin since the ampicillin-resistance gene encodes an enzyme that inactivates the antibiotic. Thus, transformed E. coli cells containing ampicillin-resistance plasmids can easily be selected simply growing the bacteria in the presence of ampicillin-only the transformed cells survive. The ara control region regulates GFP expression by the addition of arabinose, so the GFP gene can be turned on and

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.

The purpose of this experiment was to show the genetic transformation of E. coli bacteria with a plasmid that codes for Green Fluorescent Protein (GFP) and contains a gene regulatory system that confers ampicillin resistance. A plasmid is a genetic structure in a cell that can replicate independently of chromosomes. In this lab, the Green Fluorescent Protein, which is typically found in the bioluminescent jellyfish Aequorea Victoria, was cloned, purified, and moved from one organism to another with the use of pGlo plasmids. It was hypothesized that if bacteria that were transformed with +pGlo plasmids are given the gene for GFP, then transformed cell colonies

How does the addition of pGLO plasmid to a solution containing E. coli bacteria affect the growth and characteristics of the bacteria? Genetic transformation is the incorporation of foreign DNA into an organism to potentially change the organism’s trait. Plasmids are small circular DNA that replicate separately from the bacterial chromosome. In nature, these plasmids can be transferred between bacteria allowing for the sharing of beneficial genes. Due to this characteristic, plasmids allow for genetic manipulation and can be moved between bacteria easily. The pGLO plasmid utilized in this experiment encodes the gene for Green Fluorescent Protein (GFP), which under the right conditions can produce a glow. The gene regulation system present in the pGLO plasmid requires

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

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.

This experiment was completed to conduct bacterial transformation and introduce ampicillin resistant DNA plasmid to E. coli bacterial cells. Observations are made of the difference in cell growth due to various environments provided within agar

The objective of this experiment was to observe the transformation occurrence with E.coli and the ampicillin resistance gene.When plasmids like lux or pUC18 are added in E.coli, they are more likely to survive in certain environments that contain antibiotics. Plasmid can carry genes enabling bacteria like E.coli to survive in harsh conditions. This experiment displayed how plasmid work when inserted in E.coli with and without ampicillin. In certain agar plates, ampicillin was added with either the lux plasmid or the pUC18 plasmid resulting in colony growth. In other agar plates, no ampicillin was added when either the lux plasmid or the pUC18 plasmid was added resulting in colony growth. If growth occurred from the bacteria when it was added in the Ampicillin agar plates, then it was determined that the E.coli was transformed successfully and is expressing the Ampicillin resistant genes.

Biotechnology is an area of science in which nature is used for the benefit of humans. In biotechnology, scientists manipulate nature to better suit it for human application. One way that scientists manipulate nature is by genetically engineering living organisms. Genetic engineering can help humans by curing diseases and making food more nutritious among other benefits. Genetic engineering is accomplished by taking genes from one organism and planting them in another. In order to genetically engineer, different things are required depending on the organism being engineered. For example, bacteria is an organism that can be genetically engineered. To genetically engineer bacteria, scientists need bacteria, a plasmid, a stressor, an energy source, selection, and regulation. The bacteria is the nature being manipulated. A plasmid is a small DNA molecule that when inserted into a bacteria (or other cell) can change the cell’s DNA by adding to it. A stressor is an external stimulus of some type which causes a stress response. Energy is necessary for any cellular work, so it is necessary for the manipulation of bacteria. Selection shows a selectable marker, which is a favorable trait and also shows whether the DNA has been successfully planted in the bacteria. Regulation shows how a trait can be expressed. An example of a bacteria that can be genetically engineered to glow using these factors is E. coli. The plasmid in this case is pGLO, which gives the E. Coli a

The domestication of animals 30,000 years ago was the precursor to Genetic Engineering. Starting with the dog we have since been able to introduce desirable traits in all organisms. With the discovery of plasmids in the late 60’s we have been able to take genetic engineering even further. Plasmids are small circular DNA molecules used to amplify and replicate a gene of interest. These minute molecules have the ability to replicate with the chromosome or independently, allowing them to have up to a 100 copies in one cell. Plasmids are important because of their characteristics to transfer genes that occur naturally within them or acting as a vector to introduce foreign DNA into a host cell. With the use of plasmids genetic engineers are able to use bacterial transformation to make medicines. Bacterial transformation is the process in which a bacterial cell takes up foreign DNA and incorporates this DNA into its own. With the use bacterial transformation this part of Genetic Engineering has become the most important and widely used technique, creating life saving antibiotics and medicines. E.coli is generally used in these procedures because of its ability to adapt and grow exponentially. Genetic Engineers can then use large bacterial colonies, Ampicillin, and X-gal to indicate if b-glactosidase is present along with identifying the recombinant and non-recombinant colonies. The transformed bacteria that contains ampicillin will spread and survive either turning white

Genetic Engineering (GE) refers to the direct manipulation of organism DNA to alter an organism 's specific characteristic in a way for beneficial use (Nester, 2014). The host ranges from plants, bacteria, animals, and virus. The structure of the DNA includes four nitrogenous bases, including the Adenine, Thymine, Cysteine, and Thymine (Nester, 2014). The scientist isolates a plasmid from a bacteria or yeast cell, then insert the modified DNA into the plasmid. Then the plasmid is inserted into an organism DNA with the use of a restrictive enzyme to make the specific characteristic express (Nester, 2014). Through the process of cell replication, the gene will naturally multiply in future cell generation.

The bacterium being observed was E. coli, which codes for green fluorescent protein. E. coli is a bacterium that normally lives in the intestines of humans and animals alike, and can cause infection. One hopes to observe a glow from a green fluorescent protein (GFP) which is typically made by a jellyfish. This will be made possible by the additional presence of arabinose, which acts as the inducing agent. The inducing agent influences the transcription of the GFP and activates the transcriptional promoters. A promoter allows a foreign gene to fall into place much easier resulting in gene expression. It is expected that the combination of the plasmid, with the Lysogeny Broth (LB) and the arabinose (ARB) will transform the bacteria, and allow for growth of the bacteria in the presence of the antibiotic ampicillin (AMP) along with an observable glow under ultraviolet (UV) light.

This experiment was performed to test the hypothesis if LB nutrient broth, +pGLO and -pGLO Ampicillin, and Arabinose was placed in the E. coli plates, then there will be a significant growth in the newly transformed bacteria and it will possess the ability to glow under UV light. The measurements were recorded from the bent glass tube in each glass test tube. The transformation protocol tested for the newly possessed traits in E.coli bacteria. Throughout the experiment there were many probable reasons for failure. If the pipettes and sterile loop were not thrown out in between each use, a cross contamination could cause a miscalculation in the experiment causing the data results to fail. The hypothesis that was tested was validated due to the positive results with each experiment stating that newly transformed organisms due in fact pass on traits.

Agrobacterium-mediated transformation involves the transfer of a gene into the Ti (tumor-inducing) plasmid of Agrobacterium. Upon infection of the host plant, Agrobacterium exploits the immune response to incorporate its DNA into that of the host. This process results in a plant that expresses the gene of another organism that is stably integrated into the next generation [2].