Gene Cloning And Bacterial Transformation

7. We added 1.0 mol dm-3 of NaOH into tube #8 and than added 1.0 mold dm-3 of mold dm-3 HCl until a change in color is observed.

I started off the Procedure by labeling my Two tubes +pGLO and -pGLO. After that I used my pipet to put 250 ul of CaCI2 into both tubes. I placed the tubes in some ice and started to select colonies of bacteria that were "fat". I then

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

Begining by labeling 7 different 2.0 ml tubes 0 thru 6 for each compound. Then add 1ml of extract to tube 0. Then add 0.5 ml of DMSO to tubes 1 thru 6. Now make a 1:2 serial dilution from 0(pure extract) to 6(1:16)

I found that -DNA/LB was not nearly as moist as that -DNA/LB/AMP plate. The first one was dry and white.

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

It will also highlight how successfully integrated plasmids can affect the cell and what proteins are produced by the DNA, and how easy it is to change the genetics of cell and the future generations of that cell. The experiment also demonstrates how natural selection would work in a lab environment, and how the addition of a gene or a mutation can protect an organism from adverse conditions in the environment. This lab also illustrates the change in DNA in a visual sense. Molecular genetics is a very small scale science that cannot be seen by the human eye, however by illuminating the bacteria with the GFP it will produce the very small process can be clearly seen before and after the genetic splicing. The bacteria will produce quickly and the successful bacteria will be clearly seen by the human eye, even after generations upon generations of bacteria.

coli, to express a gene that is normally causes jellyfish to be fluorescent. The plasmid that contains the gene for the green fluorescent protein (GFP) is called pGLO. The type of genetic transformation we used in the experiment was heat shock which made the membrane of E. coli fluid and allowed the pGLO to enter the cell. In order for the E. coli to express the GFP gene there needs to be arabinose presence, which is a sugar that acts as an on switch for the pGLO. Using this information, we predicted the bacteria E. coli will grow and express the GFP on the plate that consists of +pGLO, ampicillin, arabinose, and LB broth (Luria Bertani broth).

Transformation is the directed modification of a genome by the external application of DNA from a cell of different genotype (Griffiths and Miller).Bacterial transformation is the easiest type of transformation to create and study due to the single cellularity of bacteria and its

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.

The heat shock treatment increases the rate of successful transformation by making the bacterial plasma membrane more permeable to the plasmid solution. During the experiment performed in the Life 102 Lab we took a sample of Escherichia Coli (E. Coli) bacteria and transformed it using a gene that codes for green fluorescent protein (GFP). (Weedman 2016). The GFP originates from a bioluminescent jellyfish, and the protein is what allows them to glow. The GFP gene is only visible in the presence of arabinose.

In the past scientist have used the bacteria Escherichia coli (E. coli) in Synthetic Biology to allow the bacterial species to recognise and respond to light frequencies providing greater knowledge of complex biological systems (Levskaya et al. 2005). E. coli is a suitable chassis to express Green Fluorescent Protein (GFP) as the bacteria’s plasmid helps to establish GFP (Chavshin et al. 2013). The bacterial plasmid is a maltose-binding protein that helps E. coli and GFP fuse together and allows for detection of gene expression and controlling internal environments (Feilmeier et al. 2000). As GFP is a useful biological indicator due to its fluorescent structure (Mazzola et al. 2005) and through the ability of E. coli responding to light, GFP is easily expressed through the process of light stimulation (Lee, J et al. 2013). Through the unique structure and functions found within E. coli, it allows for successful expression of GFP making E. coli a suitable base organism.

Genetic transformation is the process involving the insertion of new DNA into a bacteria cell such as E.Coli. While the inserted DNA contains a large chromosome, it also contains small circular pieces of DNA called plasmids. The injected plasmids hold specific genes as they are encoded through genetic engineering. Genetic engineering is described as “the direct manipulation of genes for practical purposes” (Urry et. al., 2016). The DNA injected directs the synthesis of proteins due to genetic expression and explains how bacteria is given the ability to resist antibiotics through the use of the pGLO plasmid. The “pGLO plasmid has been genetically engineered to carry the GFP gene which codes for the green fluorescent protein” (Biorad). This

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