The Importance Of Genetic Transformation

Abstract In this lab of transforming bacteria that was experiment today, I will be identifying the process of bacterial genetic transformation and how to calculate transformation efficiency. The samples that will be used in today’s bacteria will contain samples of E.coli sand inserted DNA plasmid into their genetic sequence. If done correctly the results will show a successful genotypic and phenotypic mutations, which will display fluorescent under ultra-violent lights or show signs to being resistant to ampicillin. This experiment was primarily for the purpose of growing E. Coli bacteria.

In this experiment we were meant to observe the transferring of DNA. There are many ways in which DNA can be transferred into an organism, for example; transformation, transduction, and conjugation. In our experiment we used

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

To do a transformation one micro test tube was labeled +pGLO and the other was labeled –pGLO, using a transfer pipette 250 µL of transformation solution was added to each tube in the foam rack. The tubes were then placed on ice for three minutes. During these three minutes, a sterile loop was used to pick up a single colony of E. Coli from the starter plate by gently running the loop over the agar. This loop was then inserted into the +pGLO tube and the loop was spun until the entire colony dispersed. Using a different sterile loop, the same procedure was used for the –pGLO tube. After both tubes had their own colony of E. Coli, they were placed on ice for another three minutes. DNA plasmid was added to the +pGLO tube by taking a new sterile loop and immersing it into the stock tube creating a film across the loop then inserting

This experiment focuses on genetic engineering and transformation of bacteria. The characteristics of bacteria are altered from an external source to allow them to express a new trait, in this case antibiotic resistance. In is experiment foreign DNA is inserted into Escherichia coli in order to alter its phenotype. The goal of the experiment is to transform E. coli with pGLO plasmid, which carries a gene for ampicillin resistance, and determine the transformation efficiency. The bacteria are transformed by a combination of calcium chloride and heat shock. When the bacteria are incubated on ice, the fluid cell membrane is slowed and then the heat shock

This experiment was performed to assess the efficacy of genetic transformations on bacteria via plasmid DNA coding for ampicillin resistance and green fluorescent protein. Genetic transformation was studied by taking transformed and untransformed Escherichia Coli (E. coli) and placing them on various media to observe gene expression via growth and color under UV light. The transformed E. coli were able to grow on ampicillin while the untransformed E. coli, which lacked the plasmid genes for ampicillin resistance, only grew on nutrient broth. In the presence of arabinose, the transformed E. coli glowed green. These results support the previous scientific understanding of bacterial competency, vectors, and gene expression and support gene transformations as an effective method to transfer the desirable DNA of one organism into another organism’s DNA. These results can be applied to real world issues such as medical treatments, food production, and environmental conservation.

Tiffany O’Connor                                                         November 9th, 2017              Plasmid Transformation      Purpose: Plasmid transformation is the process of transferring foreign exogenous DNA into a host cell to change its phenotype.   Theory and Background: Plasma transformation can occur naturally when a cell alters its genetics by taking in DNA from its environment. Genetic transformation can also be induced in a laboratory setting, called artificial transformation. Artificial transformation is accomplished by choosing a competent bacterium and introducing a plasmid.   E. Coli is the bacteria used in this lab.

First, 500μl of calcium chloride was added to a DNA tube. Calcium chloride makes it easier for cells to transform. Approximately 15 E. coli colonies were then transferred to the tube. 250μl of the DNA was transferred into another tube, where 10μl of pGFP was then added. The plasmid for green fluorescent protein is what will be transformed into the E. coli’s DNA so it will express the GFP. The DNA tubes were then incubated in hot and cold water .This process is called “heat shocking” and affects the permeability of cells, allowing for new DNA to move into cells more easily. Initially the tubes were put in ice for 10 minutes, they were then put in water that was 42℃ for 90 seconds, after that they were put on ice for another 2 minutes, 250μl

In transformation, a piece of DNA is inserted into a vector. Then it is cut with an enzyme and the bonded DNA insert is placed in a vector that contains DNA ligase. The insert identifies the recombinant molecules. Non-bacterial is like transformation, however non-bacterial does not require host bacteria. Phage introduction is when a phage is used instead of bacteria.

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

There are five stages to this experiment: pre-incubation, incubation, heat shock, recovery and growth, and selection. First, two micro test tubes were obtained and labeled “+” and “-”. These labels were representative of transformed (+) and untransformed (-) Escherichia coli (E. coli). 250 microliters of transformation solution (calcium chloride) was added to the test tubes and placed on ice. During the pre-incubation period a single colony was placed into the test tube labeled “+” and was completely dispersed in the solution. This was repeated for the tube labeled “-“ as well. Then the solution was returned to the ice. In the incubation period, the pGLO DNA solution was added to the tube labeled “+” but was not added to the tube labeled “-“. The tube was then returned to the ice and incubated for 10 minutes. While this was undergoing the process, four agar plated were labeled according to what they had been treated with. The plates were treated accordingly: plate 1 contained lysogenic broth (LB)/ ampicillin (amp)/ plasmids with a green fluorescent protein (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 next step was heat chock. Both test tubes were placed in a water bath that was set at 42°C for 50 seconds. After 50 seconds the test tubes were rapidly transferred back to the ice were they

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

The power of modifying genetics is at our fingertips, allowing us to change the genes of living creatures, which of course includes humans. The concern is; what genes should be modified, and which genes should be left alone. A company called 23andMe, named after the 23 pairs of chromosomes in a human cell, will provide ancestry genetic reports and uninterpreted raw genetic data using only a kit. Created in 2006, out of Mountain View, California, the company specializes in three different products or kits, an ancestry edition, a health edition, and a complete edition. Sadly, due to FDA regulations and complications, 23andMe has had to sideline the interpretation of their health-based products in the United States. Currently, 23andMe mainly

The coding region of the gene is usually fused to a promoter, most commonly used is the 35S promoter from cauliflower mosaic virus (CMV), in order to promote higher expression levels. (Snow et. al, 1997) The popular method for genetic engineering of crop plants is natural gene transfer via an Agrobacterium tumefaciens vector, a bacterium normally found in soils. The transfer-DNA (T-DNA) vector is made by inserting the desired gene fragment in between specific 25bp repeat domains in the bacterium. The vector is then inserted into the Agrobacterium and "the virulence gene products of Agrobacterium actively recognize, excise, transport, and integrate the T-DNA region into the host plant genomes." (Conner et. al, 1999) The amount of DNA transferred is only about 10kb and the nature of the gene is usually well understood. The expression of the gene introduced can also be controlled by adding additional sequences that might allow the gene to be constitutively expressed, expressed only in certain cell types, or expressed as a result of different environmental changes. This method of gene transfer, however, will only work for the natural host range of the bacterium and therefore other methods are used for additional crop plants. Such methods are uptake of naked DNA by electroporation or particle gun bombardment. The use of genetic markers, as mentioned previously, allows for the preferential growth of cultures that contain the new genetic

Instead of transferring large blocks of genes from donor plant to recipient, small isolated blocks of genes are put into the plant chromosome through biolistics, vectors, or protoplast transformation (Horsch 1993). Biolistics is a technique that shoots the gene block into the potential host cell. In order for the process to succeed, the microscopic particles and DNA must enter the cell nuclei and combine with the plant chromosome. Biolistics is commonly used but has a slight failure risk since the breeder has little control over the destination of the gene block (Mooney & Bernardi 1990). Bacteria or viruses can also carry the gene blocks into a new cell. Common vectors in gene transfer between plants are Agrobacterium tumefaciens and Agrobacterium rhizogenes. In the soil, the bacteria will infect the plants with their own plasmid, transferring the desired gene that was placed in the bacteria's DNA. Vector gene transfer is a preferred method of transformation since this modification already occurs naturally in the environment (Rudolph & McIntire 1996). Last is protoplast transformation, which uses enzymes to dissolve the cellulose in the plant wall that leaves a protoplast. Once a specific gene block is added to the protoplast, the cell wall will re-grow into a transgenic plant.