Bacteria And Bacterial Cells

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).

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

In preparing for the bacterial transformation, DNA plasmid is introduced into the E. coli cells that will express newly acquired genes. Two tubes were used and labeled both as +pGLO and -pGLO. A solution of (CaCl2) was transferred 250 µl onto the two tubes. The tubes were placed on the ice. A sterile loop was then used to gather a single colony of bacteria from a starter plate. Now, that both tubes contain bacteria they were placed on the ice for 10 minutes. Four agar plates were labeled as: +pGLO LB/amp, +pGLO LB/amp/ara, +pGLO LB, -PGLO LB/amp. Heat shock was used to transfer both the +pGLO and -pGLO, at exactly 42°C. Time was observed for 50 seconds and quickly return the tubes to the ice for another 2 minutes. As the tubes, cold down they

The acquisition of antibiotic resistance comes from horizontal gene transfer. Horizontal gene transfer refers to transferring of genes to an organism during that organisms life and not during reproduction. This acquisition comes from transduction, transformation and conjugation

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

It was also conducted to learn about the process of moving genes from one organism to another with the help of a plasmid. The control group was the -pGlo with LB and ampicillin antibiotic, and the -pGlo with LB. The experimental group was the +pGlo with LB and ampicillin antibiotic, and the +pGlo with LB, ampicillin antibiotic, and arabinose. The dependent variable was the bacteria. It had the ability to change. The independent variable is the ampicillin antibiotic and pGlo. They stand alone and are not changed by other factors. The organism used was the bacteria GIVE TYPE OF BAC. and HOW WILL THE BACTERIA BE

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].

the bacterial chromosome is genetically engineered and the plasmid is used to help the bacterium replicate.

Experiments introduce the idea that the uptake of DNA by competent bacteria can be the result for survival reasons such as need for food or evolution purposes (5). The difference between natural and artificial gene transformation in a bacteria, however, with those who are incompetent versus competent allow he or she to conduct an experiment with artificial transformation. By doing this one may be able to determine and observe the formation of biofilms or mutations by transferring a gene to another organism for beneficial or harmful purpose. This could lead to finding resistants or if a gene introduced has any effect on the bacteria. In the experiment, the pGLO plasmid contained encodes the gene for GFP as well as a gene for resistance to an antibiotic allowing a transformation to take place when adding a carbohydrate such as arabinose to the medium. Research has been found on the horizontal gene transfer on how virulence factors are acquired as well as genes resistant to antibiotics spread with microorganisms (5). By conducting a similar experiment he or she can observe similar findings or observe the results and conduct some sort of understanding of these ideas. The findings in this experiment are to show a positive result in what causes growth and the glowing when the plasmid pGLO is added to the positive as well as the difference in the medium. Understanding the way artificial

Abstract:Conjugation is a natural occurring process that involves the transfer of DNA from one cell into another through a physical connection between the cells. In the following experiment, two strains of Escherichia coli bacterial cells (donor F'lac+strs and recipient F-lac-strr) underwent conjugation to produce a transconjugant strain (F'lac+strr). MAC plates and streptomycin were utilized to determine if conjugation had occurred. When plated, the donor colonies appeared red and the recipient colonies appeared white. The transconjugant plates showed red and white colonies. Using alkaline lysis miniprep, a DNA plasmid was isolated from the donor and transconjugant strains and FIGE electrophoresis was used to determine the size of the

Bacteria are microorganisms that are well known to be the major cause of why people get ill. Many of these microorganisms in the world are pathogenic by nature but are treatable by a wide number of antibiotics, some are normal flora in the body, and others are opportunistic ones. In recent years, scientists formulated specific antibiotics that would eliminate and eradicate these pathogenic types of microorganisms, but there are still some microorganisms that are present today that is hard to eliminate due their capability to tolerate antibiotics.

Bacteria have evolved so that they can transfer different strains of resistance to their own species and different species (Khachatourians, 1998). The organisms have genetic traits in the chromosomes and extrachromosomal elements for resistance to occur (Khachatourians, 1998). Resistance can occur from a mutation on the chromosomal genes, leading to antibiotic sensitivity. Khachatourians states that, “such mutations occur at a rate of one per million to one per billion cells”. He also explains that, “the extrachromosomal elements (plasmids and transposons) are smaller pieces of circular DNA, each equivalent in size to about 1% of a chromosome” (Khachatourians, 1998). When the bacterial organism acquires the resistance against the antibiotic administered and send it to other species of bacteria this is known as genetic exchange (Khachatourians, 1998).

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

Biological science has progressed rapidly in the last two centuries and nowadays, more than ever before, human beings have the capability to change their environment. One of the ways we do it is genetic engineering. It may be observed that a lot of products have the label GMO (Genetically Modified Organisms). These products are manufactured using genetically modified plants. These plants usually keep their non-modified form, but change their reaction toward the environment. Genetic modification is used to protect them from unwanted consequences, such as being eaten by a predator. It is an advantage in the production, but since a human being has to consume it, the change in DNA (Deoxyribonucleic Acid) of the plant may have an effect on our organism.

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.