Personal Narrative: My Best Bacterial Gene

This lab is about moving genes from one thing to another using plasmids. Plasmid has the ability to replicate, so it replicates independently, and separately from the chromosomal DNA. Plasmid are one or more small piece of DNA and they enter cells as a double strand DNA. When they enter the cell as a doubke strand they do not invade he chromosomal DNA. We will also transform bacteria into GFP which is mainly from the jelly fish Aequorea Victoria. The GFP causes the the jelly fish to fluorescent and glow in the dark. After the transformation, bacteria starts to make the GFP which causes them to glow a green color under a ultraviolet light.

134). They are loops of DNA that are separate from the chromosomal DNA and can self-replicate in a cell, found mostly in bacteria (Brown, 2011; Addgene, 2015). Lederberg and William Hayes discovered that plasmids were being transferred from one cell to another, not the chromosomal DNA (Brown, 2011, p. 135). This discovery lead to plasmids being an essential tool for scientists. Scientists can engineer plasmids to have specific genes to introduce into new cells (Brown, 2011, p. 134). On a plasmid loop there will be an origin of replication (ORI) and a multiple cloning site (MCS) where the gene of interest is inserted (Bio-Rad, 2015). This region has specific restriction enzyme recognition sites, which are cut by the enzymes to open up the DNA where the new gene will be inserted (Jove Science Education Database, 2015). Most plasmids will also contain an antibiotic resistance gene allowing cell survival in environments containing antibiotics (Jove Science Education Database, 2015).

This experiment was designed to test and observe the transformation efficacy of the pUC18 and lux plasmids in making E. coli resistant to ampicillin. Both plasmids code for ampicillin resistance, however, the lux plasmid codes for a bioluminescence gene that is expressed if properly introduced into the bacteria’s genome. The E. coli cultures were mixed with a calcium chloride solution and then heat shocked, allowing the plasmids to enter the bacteria and assimilate into the bacterial DNA. The plasmids and the bacteria were then mixed in different test tubes and then evenly spread onto petri dishes using a bacterial spreader, heating the spreader between each sample to make sure there is no cross contamination. Each of the dishes was labeled and then incubated for a period of 24 hours. The results were rather odd because every single one of the samples grew. Several errors could have occurred here, cross contamination or possibly an error in preparation as every single sample in the class grew, meaning all samples of the bacteria transformed and became ampicillin resistant.

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

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

Certain E. Coli strains are also known to show resistance to bacteria killing antibiotics. This resistance is due to the plasmids, or small round DNA molecules, in the bacteria that carry the resistant genes. R Plasmids (resistance plasmids) are widely studied and bestow resistance to factors that inhibit growth of the organism. R plasmids code for proteins that can

As I watched my mom cultivate relationships that were real and substancial to her well being I quickly realized that these were also the kind of relationships that I wanted. I think that the reason these relationships where people are real can be so growing for both sides, being real with people can be hard sometimes, but in the end both people can come out better. As I look at the life that Jesus lead I am convinced that Jesus was the kind of "keep it real" guy. A prime example is looking at him the night before he was crucifed, he sat in the garden and asked that if at all possible that God would take away what he knew was coming. That is real, and that shows us how he was human. There are so many other examples how Jesus' life was

Juxstapositions of “The Stolen Bacillus” and “The Adventure of the Bruce-Partington Plans” The Scientific Method is a research tool used to analyze the realities of life by exploring new phenomena, aquiring knowledge about the world around us and then, developing that data into questions that will open up to further investigation. Moreover, this method made up of theories/data that lead to the formation of a hypothesis. The hypothesis is the most important part of the experiment because it constitutes inductive and deductive reasoning, but more importantly it is falsifiable and realistic. This leads to the experimental portion of the scientific method where the researcher will test his/her predictions and draw a conclusion based on the given

When antibiotic is used most of the bacteria die but a few bacteria with antibiotic resistance gene survive and reproduce and pass this advantage to their offsprings. This selective pressure exists naturally, however antibiotic misuse can be accused for fastening the spread of the antibiotic resistance gene [Refer to figure 2] (Learn Genetics 2015). Consequently, inappropriate antibiotic intake will lead to a greater chance of superbugs being developed. Antibiotic resistance can be defined as a new ability which a bacterium has developed to stay unattached in the presence of an antibiotic that was previously effective to destroy the bacterium (ABC science 2015). Four key mechanisms that has been identified for bacterial antibiotic resistance can be listed as: producing enzymes that inhibit the functionality of the drug, reducing the effectiveness of the drug by producing targets against which the antibiotic, reducing the permeability of the drug into the bacterium and active export of antibiotics using various pumps (Pogson 2012). All these mechanisms can be developed by any of the bacteria when the corresponding mutated gene of antibacterial resistance is received. The genes code for specific proteins, and variation in the gene leads to alteration of the shape of proteins. This leads to changing the functionality

Escherichia coli is a well studied prokaryote and may be considered a foundation of biotechnology with the work of Herbert Boyer and Stanley Norman Cohen who used E. coli plasmids in conjunction with restriction enzymes and produced recombinant DNA.1 Even before the application of the rDNA technology, the first industrial application of E. coli being the production of the amino acid threonine in 1961 by the induced mutagenesis of the

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

Plasmids are small double stranded circular non chromosomal DNA molecules containing their own origin of replication. Hence, they are capable of replication independent of the chromosomal DNA in bacteria. Plasmids present in one or more copies per cell, can carry extra chromosomal DNA from one cell to another cell and serve as tools to clone and manipulate genes. Plasmids used exclusively for this purpose are known as vectors. The genes of interest can be inserted into these vector plasmids creating a recombinant plasmid. Recombinant plasmids can play a significant role in gene therapy, DNA vaccination, and drug delivery [Rapley, 2000].

Overtime, the evolution of bacteria has been utilized as a tool to produce medicines such as insulin and even varios growth hormones. Transformational success of bacteria is mainly due to versatile genetic coding, enabling researchers to transfer DNA to numerous amounts of organisms. Studies have utilized quantitative analysis to design experiments and expose Escherichia coli, commonly known as E-Coli, to a plasmid surface obtaining genes that have the potential to resist Ampicillin, which is identified as an antibiotic. A substance known as Calcium chloride was used to eliminate repulsion that existed between the DNA and cell wall of the bacteria. The Heat shock method was utilized to give DNA a required energy boost to enter the bacteria

Did you know that with the science of DNA manipulation, animal cadavers can be turned into insulin for diabetics? Back in the 80's scientists isolated the human gene for insulin and transferred it into bacteria. Now bacteria cultures are used to produce large amounts of human insulin. DNA manipulation is especially important in medicine, where it holds the hope of curing genetic diseases such as Huntington's, and even some types of cancer. There are several major types of DNA-manipulation enzymes used by living cells. The first type is DNA polymerase, which cells use to replicate their own DNA. The next type is DNA ligase, which joins 2 pieces of DNA to create a single piece. The third type is restriction enzymes, which appear to be made only by bacteria. Restriction enzymes are very important enzymes that are vital to our manipulation of DNA (Rapoza, DNA, 2).

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