Heritable Genetic Information In Escherichia Coli In Transformation

Models provide the physical testing and proof of a hypothesis by exploring the extent to which the two factors relate within the given hypothesis. It puts a theory into action, to see if the theory is corrected causes and effects.

The results of this experiment ended up being very conclusive and not positively or negatively surprising. The data and results collected we what we had expected more importantly the ones on the X-Gal plates where the colonies were above 100. Both the stains of E. Coli grew colonies when they were in the LB. When the ampicillin was present the cells did not grow even with the X-gal. The transformed cells on the other hand grew when the ampicillin was there. When the X-gal and AMP were present the transformed cells grew and had a blue tint, which is why we call it the pBlu lab. (duh)

Models provide the physical testing and proof of a hypothesis by exploring the extent to which the two factors relate within the given hypothesis. It puts a theory into action, to see if the theory is correct.

As predicted the E. coli colony transformed with either the PUC18 or the lux plasmid developed an ampicillin resistance. Which made it easier for them to not only survive but also replicate in both the LB agar plates and the LB ampicillin rich agar plate. However the E. coli colony not treated with the plasmids could not survive and colonize in the LB ampicillin rich agar plates. The plate that had no ampicillin in its environment and no plasmid treated E. coli served as a positive control for this experiment because it demonstrated how the E. coli would colonize and grow in a normal setting. The cells in the positive control plate grew into lawn colonies because they were not placed into a selective environment or transformed, so they had no need to acquire ampicillin resistance. Two plates in the experiment contained E. coli cells that were transformed with either the PUC18 or the lux plasmid but were placed in an ampicillin free environment. These two colonies grew

E. coli HB101 was transformed with pGLO plasmid then grown on media containing ampicillin and/or arabinose and on medium containing neither (Brown, 2011). This is done for selection of transformed cells since not all cells are expected to take up the plasmid (Brown, 2011). We also expect roughly the same CFU on any plate(s) receiving samples from the same microcentrifuge tube, since they are getting the exact same

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

70µL of competent E.coli are added to both test tubes; pUC18 and Lux (Alberte et al., 2012). Both test tubes are then tapped and placed back into the ice bath for 15 minutes. While waiting, another test tube is obtained, filled with 35µL of competent cells and labeled NP for no plasmid. A water bath is preheated to 37 degrees Celsius and all three labeled test tubes are inserted into the bath for five minutes (Alberte et al., 2012). Using a sterile pipet 300µL of nutrient broth are inserted into both the control and Lux test tubes and 150µL are inserted to the no plasmid test tube to increase bacterial growth. All three test tubes are then incubated at 37 degrees for 45 minutes. Six agar plates are obtained and labeled to correspond each test tube, three of the plates contain ampicillin. A pipet is used to remove 130µl from each test tube containing a plasmid and insert it into the corresponding agar plate. For this, a cell spreader is first

Escherichia Coli, or E. Coli, can also form a number of these substances itself with the use of energy. (Shiloach and Rinas, 2009b) Understanding that it can make these substances with energy, E. Coli can reproduce asexually to make identical cells through binary fission. (Souza, Castillo, and Eguiarte, 2002. )

Instead, they argued in favor of the recombination dependent mutation theory. E.coli with a Lac- mutation were found to revert back into Lac+ bacteria when grown in lactose and minimal media. However, the mechanism was still unclear. Some scientists argued the lac region still had some ability to catalyze lactose, and will be amplified until it can grow on lactose. On the other hand, scientist at Indiana University state revertants are created through DNA error. Through three experiments, scientist argued against the amplification dependent mutation theory. Transposons were inserted at various distances from the lac region, and transformation frequency did not decrease as the transposon and lac distance decreased. Amplification of the transposon and the lac region should have occurred at the same frequency because they are close together on the plasmid. Furthermore, tetA gene, another gene on the plasmid, was also selected in the experiment. Similarly, the reversion of this gene was not tied to the reversion of the Lac- no matter the distance. Lastly, the amplification theory states alleles must be cis to undergo amplification. However, scientists found that reversion still occurs when the lac region is trans. Thus, these scientists debunk the amplification dependent mutation theory in favor of the recombination dependent mutation

Escherichia coli, or E. coli, is a common bacterium that can be found in diverse environments all over the planet, including the gastrointestinal tracts of animals and humans. Many of these strains of E. Coli are essential mechanisms in the digestive tract, while others are pathogens that can cause complications in urinary and intestinal tracts. (Payne & Sparks) In research, E. Coli is commonly used as a model organism, meaning they are widely studied by scientists for a variety of purposes due to their experimental advantages. E. Coli is comparatively simple, and there are many advantages to studying these prokaryotic cells in the fields of biochemistry and molecular biology. E. Coli has this simplicity and is relatively easy to propagate in a lab environment. Their genome has been completely sequenced and many things we know about DNA, protein synthesis, and gene linkage have been derived from studies regarding this particular organism. (Cooper)

If a gene that codes for Green Fluorescent Protein transforms bacteria and GFP glows when transformation occurs, then when two micro test tubes have 250 microliters of transformation solution and places in an ice bath, then 2-4 bacteria colonies are added to each tube with a sterile loop; then a plasmid (pGLO) is added to one of the tubes, incubated in ice for 10 minutes, then heat shocked for 50 seconds at 42 degrees Celsius, then back into 9ice for two minutes; then LB nutrient broth is added to both tubes (250 microliters) and set out at room temperature for 10 minutes. Then, 100 microliters of each solution in the tube are added to four

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

The first step of this experiment was to determine if genetic transformation has occurred within the E. coli culture. The plates that were compared in this section can be seen in Figure I. The plates that were compared were labeled LB+ amp (+) and LB + amp (-). The positive control was the untransformed cell in LB+ amp(-). This control was testing: if the antibiotic would kill all of the untransformed cells in the LB+ amp (+). In figure I. the plate labeled LB + amp (-) had no bacteria on it. Therefore, the antibiotic did kill all the bacteria that did not transform. In conclusion, the plate labeled LB+ amp (+) contained only transformed cells, and this can be seen In figure I., because the bacteria on the plate labeled LB+ amp (+), are

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