Bacteria will cultivate in the presence of pGLO, and the student whose sample contains arabinose will exhibit fluorescence under the UV light even after DNA isolation, quantification, restriction digest, and protein analysis.
Discussion/Analysis.
The experiments begin with the transformation of pGLO plasmid in E.coli. We observed growth in each of the three cultures due to the presence of the LB broth and the pGLO. Any bacteria present on the LB/AMP and LB/AMP/ARA plates exhibit successful bacterial transformation, because only the bacteria with the transformed DNA can grow in the presence of ampicillin. The third plate; Figure 3, (LB/AMP/ARA) glows under the UV light due to the presence of arabinose (ARA), it activates the expression of the GFP gene that contributes to fluorescence. .
For the next lab, the transformed DNA is isolated, and then centrifugation is used to collect a cell pellet to be used in later labs. I had the culture with ampicillin and arabinose, while Mohammed's culture contained no arabinose. Under the UV light, my pellet glowed and his did not. This observation is due to the arabinose in my pellet. .
The subsequent class involved quantification and restriction digest. After diluting our samples, we measured absorbances at 260 nm and 280 nm. My readings were 0.482 and 0.273 respectively, and the quality of my DNA was 1.76. Mohammed obtained a similar value for his sample (1.79). The DNA concentration for the crude prep is usually not more 1 mg/ml (micro), and any numbers greater than that usually indicates contamination. In our case, the DNA quality reading is expected to be greater than 1 because we have RNA in our samples, which also absorbs at a λ max of 260 nm. The reason why there is RNA in our samples is because we did not use any RNAse or other enzymes to degrade it. .
The following week involved the use of electrophoresis to separate sample components and isolated the GFP protein.
Ofloxacin) prevent bacterial growth by hindering the activities of bacterial DNA Gyrase and Topisomerase. Sulphonamides slow bacterial growth by inhibiting specific bacterial metabolic pathways. ... The secondary method of resistance is transformation. In transformation, bacteria intake segments of DNA and incorporate it into their own DNA. ... These bacteria have an increased capacity to uptake DNA. ...
Debates over whether it was proteins or DNA that were the bearers of "genetic instruction" for human life, (even though DNA was located exclusively ON the chromosomes), kept the latter out of the picture until 1944 (DNA Watson). ... Based on the work of a British scientist (in which a heat-treated form of Pneumococcus and an innocuous strain of the same bacteria combined to return the first to it's lethal state), Avery et al. systematically destroyed individual parts of the bacteria's machinery through the use of enzymes to determine what made the transformation possible (DNA Watson...
One important application regarding genetic engineering is to combine the DNA from various organisms into bacterial cells to produce a required product. ... It is the act of taking a piece of DNA and combining it with another organism's DNA to produce a new strand of DNA. ... Once the donor DNA has been chosen, a vector must be used to implant that DNA into the bacterium. ... This newly formed DNA is recombinant DNA. The recombinant DNA formed after the donor DNA and the vector DNA were combined must now be introduced into the bacteria, and this process which allows the bacteria to be pla...
During attachment, portions of the capsid combine with a receptor on the rigid bacterial cell wall in the lock-and-key manner. During penetration, a viral enzyme digest away part of the cell wall, and viral DNA is injected into the bacterial cell. ... Many bacteria have extra chromosomal rings of DNA called plasmids, which are often extracted and used as vectors to carry foreign DNA into other bacteria during genetic engineering. ... Sexual exchange of DNA occurs among bacteria in three ways. ... Transformation occurs when a bacterium binds to and then takes up DNA released into the medium...
The transformation time from tree-shrew, to ape, to human far exceeds the time from analytical engine, to calculator, to computer. ... It will usher in a world where gene defects, bacterial disease, and even aging are a thing of the past. ... "Transforming bacterial cells is easier than transforming the cells of complex organisms" (Stableford 34). ... This now indestructible bacterial pestilence wages havoc on the human body. Genetic engineering is conquering this medical dilemma by utilizing diseases that target bacterial organisms. ...
The transformation time from ape, to human far exceeds the time from an abbacus, to a computer. ... In essence, scientists aim to remove one gene from an organism's DNA, and place it into the DNA of another organism. ... The restriction enzymes cut the DNA strand at a particular location called a nucleotide base, which makes up a DNA molecule. Now that the desired portion of the DNA is cut out, it can be joined to anothe strand of DNA by using enzymes called ligases. ... This can be accomplished by using special pieces of DNA, called vectors, that permit the generation of multiple copies ...
Plasmid is a small ring of DNA that carries accessory genes separate from those of bacterial chromosome. ... In nature genes can be transferred between bacteria in different ways like transformation, conjugation, and transduction. One-way is bacterial transformation, it is when transfer of genetic information into a cell by the direct uptake of the DNA. Then the DNA is used to transform the cells from the original DNA to take a certain trait. How ever these bacteria can take up DNA only during the period at the end of logarithmic growth. ...
DNA copies itself by semi conservative replication. This means that each new molecule of DNA contains one strand from the original DNA and one newly synthesized strand. ... The bacteria incorporated the N15 in their DNA, making the DNA denser than normal. ... This DNA may then be incorporated into the bacterial DNA, known as transformation. ... When this phage infects another cell, the acquired gene is inserted into the bacterial genome. ...
Now scientists know much more of genetics and DNA. ... Some indirect evidence showed DNA to be related in some way to genetic material. ... DNA was not, however, at this time considered to be identical to genetic material, mostly because some chemical test had shown that DNA probably did not have the chemical diversity needed for a genetic substance. ... This ability allows it to be immune to the attacks upon it by the bacterial defenses in the body of the infected animal. ... This experiment showed that DNA was the chemical that perpetrated the genetic transformation, ...