discovery and characterization of restriction enzymes first took place in the late 1960’s and early 1970’s. The scientists responsible for the discovery were molecular biologists Werner Arber, Hamilton Smith and Daniel Nathans. In the late 60’s Arber observed a sharp change in the bacteriophage DNA he had been working with after it invaded resistant strains of bacteria. It had been cut into pieces and degraded. He hypothesized that bacteria could express two different enzymes: one that recognizes and destroys
Genetic engineering is possible because of special enzymes that cut DNA. These enzymes are called restriction enzymes or restriction endonucleases. Restriction enzymes are proteins produced by bacteria to prevent or restrict invasion by foreign DNA. They act as DNA scissors, cutting the foreign DNA into pieces so that it cannot function. A nuclease is any enzyme that cuts the phosphodiester bonds of the DNA backbone, and an endonuclease is an enzyme that cuts somewhere within a DNA molecule. In contrast
The pK184hlyA and pBluescript plasmids were cut using EcoR1 and Pst1 restriction enzymes that cut at specific restriction sites. Gel electrophoresis was carried out to check if the digestion procedure was done successfully. Figure 2 shows the results of the electrophoresis. Lanes 5 and 7 indicate the fragments obtained when the plasmids are digested with both restriction enzymes, indicating the approximate fragment size for the hlyA gene, the pK184 plasmid and the pBluescript plasmid. This is useful
Once the plasmids are cut in a restriction digest reaction, it is then run on an agarose gel. It is important that this is done. The digest gel is used as a confirmation test. It allows one to verify whether or not the restriction enzymes cut the plasmids at a specific sequence; by comparing the base pairs of the product to the 1 Kb ladder (provided one knows the number of base pairs in the expected fragments). One will be able to determine whether the plasmid cut, and if it cut the fragment like
This experiment was very successful as a credible restriction map for the unknown plasmid could be constructed. Within this experiment, both single digest and double digests consisting of three restriction endonucleases were used in order to map out the restriction sites of the enzymes making up an unknown plasmid. In order to separate the DNA fragments by their distinct number of base pairs, it was necessary to run an agarose gel electrophoresis. Within the gel electrophoresis, it is necessary to
Restriction enzymes and DNA ligase are very important parts in theprocess of molecular cloning. In this experiment, Lambda DNA was digested using restriction enzyme HindIII and then ligated back together using DNA ligase. The partially ligated DNA and the fully ligated DNA were compared to the original DNA side by side on an agarose gel. The results showed that a significant amount of Lambda DNA was digested and the ligation stages had visible progress. Introduction Restriction enzymes (restriction
plating the transformation cells onto LA plates that have either ampicillin or chloramphenicol and PCR. We will use the PCR thermocycler to denature the restriction enzymes that were specifically used to assimilate the vector DNA. It is important to use the PCR thermocycler because denaturation of the restriction enzyme will prevent the restriction enzyme from cutting the vector DNA, after the insert DNA has assimilated to the vector DNA. After the addition of specific primers that complement the base
treated with restriction enzymes, which cut the DNA at their specific sites. Purpose The purpose of this lab was to understand how to analyze a series of genetic data from DNA using agarose gel electrophoresis and decode the results. Materials Micropipette Micropipette tips Premade Agarose gel Gel electrophoresis machine DNA samples ‘Marker’ sample UV light box Methods 1. 1% agarose gel was made and allowed to solidify, and put in electrophoresis. 2. DNA was incubated with restriction endonuclease
Background Each year snakes envenomate 421,000 people, 20,000 of whom die. These injuries are especially concentrated in developing countries, where snake bites are an occupational hazard. (Kasturiratne et al. 2008). The negative impact of this could be alleviated by the creation and production of a low-cost, human-compatible universal antivenom. Lethal Toxin Neutralizing Factor, henceforth LTNF, is a substance that has been isolated from opossum (Didelphis virginiana) serum, liquid component
Endonuclease V-DNA Repair Enzyme Endonuclease V also known as EndoV is a highly conserved DNA repair enzyme. According to the research done in past, this enzyme was first discovered in E-coli and then later homologs of this enzyme were also found in prokaryotes, humans, and eukaryotes. This biochemical analysis of this protein is still not well understood by the scientist; however, Thermotoga maritime (Tma) which is a thermophillic bacterium is used as a great model to explain its enzymatic properties
that passes on the genetic characteristics from one generation to the next. DNA is found in the chromosomes in the nucleus of a cell. "Every family line has it’s own unique pattern of restriction-enzyme DNA fragments. This variation in patterns of DNA fragments found in human genetic lineages is called ‘restriction-fragment length polymorphism’(RFLP)." (Louis Levine, ?) Because each person, except for identical twins(which have the exact same DNA), is formed from two family lines the pattern of sizes
A Psychoanalytic Interpretation of Rapunzel The familiar story of Rapunzel, as told by the brothers Jacob Ludwig Carl and Wilhelm Carl Grimm, takes on new meaning with a psychoanalytic interpretation. It is a complex tale about desire, achievement, and loss. The trio of husband, wife, and witch function as the ego, id, and superego respectively to govern behavior regarding a beautiful object of desire, especially when a prince discovers this object. The story begins in a rural house where
finds that a life of activity and contemplation satisfies our purpose, achieving the most complete happiness in us. Aristotle is correct regarding the necessity of activity, but restricts the theory to only the life of study. We will reject this restriction, and instead allow any life of virtue and productivity to substitute for Aristotle’s life of study. One primary means of remaining active to achieve happiness includes loving friendships, which only happen to the virtuous. Thus human flourishing
James to speak to present-day philosophers as well as his turn of the century audience, then this reading of the Sophistes will clarify the current philosophical temperament. Neither James nor the contemporary philosopher is satisfied with any restriction on All; for this reason, both lack interest in being. Being, once the richest word, no longer satisfies the philosopher's greedy temperament. Introduction In the first chapter of Pragmatism (2) William James speaks about a rather unusual aspect
research in the Microsoft Network using the Custom Search feature in the Stock Screener. Since I was looking for a conservative stock to start with, the first restriction I put was that the stock be a member of the Dow Jones Industrial Average. I also wanted a Large-Cap company so I put the Market Cap to be above 5 billion dollars. Another restriction I added was that its P/E ratio be the lowest possible and that the EPS Year to Year be above 13%. After running the search I got 20 matches. The top three
relatively few poems. Milton was greatly influenced by the puritan cause and Oliver Cromwell with a strong parliament government. Pope was born shortly after Milton's death and was a Roman Catholic. Many restrictions against Catholics. Pope had to struggle for position. Some of the restrictions made Pope move outside London and he could not legally vote, hold office, or attend university. Pope not allowed to attend university would be one of the most significant contrasts between Milton and
Driving Restrictions There are many accidents everyday that are costing people their lives. My young brothers and most definitely my grandparents would probably kill me for saying so, but there should be some changes made to the laws concerning the age requirements of drivers. If the State of Ohio changed the age requirement of operating a vehicle to between 18-70 years of age, our roads would be much safer to drive on. If you had told me when I turned sixteen that I couldn’t get my license until
persisted as an effective cheap means of replicating DNA of interest in a foreign host cell. “To "clone a gene," a DNA fragment containing the gene of interest is isolated from chromosomal DNA using restriction enzymes and then united with a plasmid that has been cut with the same restriction enzymes.”(Human Genome Project) Plasmids are not part of the chromosome but they replicate along with the cell when it replicates and divides. Since they are not a part of the chromosome they are easier to isolate
parameters of the terms in your essay? There are two important elements here. One: “definitions” meaning that you define the words such that you can write your essay. Two: “parameters” meaning the things that you are not going to talk about or the restrictions of your essay. The Body The body of the essay comes next and many people actually don't know how to do this properly. They actually write whatever they want and whatever comes to their mind. This is incorrect. Here's how you actually do write
In the best-selling novel entitled Memoirs of a Geisha by Arthur Golden, Golden examines the secret world of the geisha. Contrary of what is often believed, geisha are far from being prostitutes; they are more accurately High-class Japanese entertainers. Arthur Golden shows the reader a completely different look on life in looking into the lives of geisha in mid-twentieth century Gion and sends a very strong message distinguishing the geisha and the prostitutes. Arthur Golden, throughout Memoirs