Dna And Multiple Cloning Sites Essay - 1407 Words

Introduction In this investigation pUC19 plasmids were used as the vector due to its small size of 2686bp, high uptake efficiency by the host and fast replication time. Important features of this plasmid include the origin of replication and multiple cloning sites (MCS). The origin of replication allows the plasmid to replicate inside the host bacterium. The MCS is located within the lacZ gene and contains unique sites for the Xbal EcoRI restirction enzymes to cut and produce sticky ends for the CIH-1 gene to bind to. Furthermore, the pUC19 plasmid also contains an ampiccilin resistance gene so only transforemed E.coli are able to remain viable when spread on the agar plates that also has the addition of ampiccilin. The lacZ gene encodes the ÃŽ ²-galactosidase enzyme which aids in indentifying the recombinant E.coli from the non recombinant cells (Coventry University 2016). The addition of 5-bromo-4-chloro-3-indoyl-ÃŽ ²-thiogalactoside (X-gal) in the agar mix acts as an artifical substrate for the enzyme so it produces blue E.coli colonies when hydrolised by the ÃŽ ²-galactosidase meaning those specific colonies do not contain the plasmid with the CIH-1 insert (non recombinant E.coli) (Coventry University 2016). Isopropyl-ÃŽ ²-D-thiogalactoside (IPTG) an artifical transcription inducer of the lacZ gene is also added into the agar mix, binding to the repressor gene and inactivating it. Therefore when the CIH-1 gene is incorporated within the plasmid MCS and inserted into the E.coliShow MoreRelatedPolymerase Chain Reaction Essay1003 Words   |  5 PagesOne may view cloning as copying a living thing and producing multiple copies. People may think of cloning rabbits, sheep or humans. In the field of molecular biology, however cloning is viewed at a genetic molecular level, where a piece of DNA is copied on a large-scale by genetically copying tens to hu ndreds of thousands of identical DNA fragments. Researchers are developing new methods of cloning by using polymerase chain reaction (PCR). PCR was introduced in the 1980s and in recent years KaryRead MoreEscherichia Coli : Is A Well Studied Gram Negative Bacteria916 Words   |  4 Pageslevel of DNA supercoiling, an important property of DNA and chromatin (Gilbert and Allan, 2014). The supercoiling level is modified by a reaction that consists in the transient breakage of the DNA phosphodiester bonds and the movement of strands across the transient breaks (Tse-Dinh and Wang, 1986). In prokaryotes, these enzymes can be classified into two major groups, according to the mechanism of action. While the topoisomerase II (DNA gyrase) generates negative supercoils into relaxed DNA and relaxRead MoreDna Lab Report1345 Words   |  6 Pages The genomic DNA sample concentration of 28.5 ng/ÃŽ ¼L and its A260/A280 ratio of 1.85 indicates that the DNA was relatively pure since a 260/280 ratio of ~1.8 is generally accepted as pure for DNA (Cox, Doudna O’Donnell, 2015). The ratio of white colonies to blue colonies observed on the blue-white screen plate was 31:37, which shows that there was a greater number of E. coli DH5-ÃŽ ± that did not contain the plasmid with the foreign gene insert. Therefore, out of all of the E. coli DH5-ÃŽ ± cells thatRead MoreVibrio Fischeri Is A Bacterium That Served As The Model Organism802 Words   |  4 Pagesexperiment, Vibrio fischeri chromosomal DNA was isolated by lysing the bacterial membranes and removing the proteins and lipids. Addition of a lysozyme solution breaks down the peptidoglycan layer in Gram-negative bacteria and proteinase K/SDS degrades the proteins and disrupt the membranes. Liquid-liquid extraction using Tris-buffered phenol permits separation of proteins and lipi ds from the DNA into the organic layer. It is also essential that the isolated DNA is pure and free of proteins, sugarsRead MoreA Segment Of The Will Die Slowly Gene From Drosophila Melanogaster1688 Words   |  7 PagesTitle: â€Å"PCR amplification, TA cloning, sequencing and sequence analysis of a segment of the will die slowly gene from Drosophila melanogaster.† Abstact: A seemingly novel gene sequence has been discovered in fruit flies from Nicholas Harden’s lab at Simon Fraser University using random PCR primers. Attempts at purifying and characterizing this gene have been elusive. Here, we show methods for isolating, amplifying, and purifying the gene of interest for analysis. Using polymerase chain reactionRead MoreEssay On Mutation1657 Words   |  7 Pagesframeshift mutations. How are these mutations the same and how are they different? Silent, missense, and nonsense mutations all occur when base substitutions happen (text p.206). A base substitution is when the wrong nucleotide is incorporated during DNA synthesis, this is the most common mutation (lecture). In a silent mutation a base substitution occurs, incorporating an incorrect nucleotide, the result is a codon that codes for an amino acid that is the same as the wild type (text p. 206). In a missenseRead MoreVectors Used in Biotechnology2739 Words   |  11 Pagesclosed circula r, double-stranded, extrachromosomal DNA molecules which occur naturally in bacteria, yeast, and some higher eukaryotic cells, and exist in a parasitic or symbiotic relationship with their host cell (Lodish et al., 2000) The main application of plasmids is as cloning vectors in gene cloning. In gene cloning, a fragment of DNA, containing the gene to be cloned is inserted into a circular molecule called the â€Å"vector† to produce recombinant DNA molecule. Plasmids are one of the most commonlyRead MoreGene Regulation For Safety Assessment946 Words   |  4 Pagesapplied to all procedures, and universal precautions and lab safety protocols are followed. Transformation Methods: 1. The Isolation of Gene of Interest from Pseudomonas putida 2. The Cloning Process 3. Transformation of bacteria Quality Control: 1. Selection and screening of Recombinant cells 2. Purification of DNA 3. Purification and Storage of Bacteria. 1. The Isolation of Gene of Interest from Pseudomonas putida: The isolation of P.putida strain was carried out by using P-2 media. This mediaRead MoreStem Cell Research1167 Words   |  5 Pages. Stem cell is a cell that has the ability to change into several or multiple types of cells or into any cell type in the body. There are two main types of stem cells, adult stem cells and embryonic stem cells. Embryonic stem cells are found in cells that have only undergone the first few dozen cycles of growth and division following the fusion of egg and sperm. These cells are pluripotent and can be maintained so that they grow and divide indefinitely without differentiating. Likewise, theyRead MoreDescription And Function Of Mutagenesis1345 Words   |  6 PagesSite-directed mutagenesis is a method used to construct amino acid changes in a protein in order to test the function of a specific amino acid. It is the way to study protein structure and function change the amino acid sequence of a protein by altering the DNA sequence of cloned gene4. A particular amino acid is very important in catalytic activity, ligand binding, protein folding or other function. Amino acid residue significance is tested by making conservative substitutions or by changing the

The Effects Of Excessive Unhealthy Food Intake - 1327 Words

One of America’s top leading problems in the 21st century is caused by vast amounts of unhealthy food which leads to not only obesity, but other health related issues such as diabetes and other heart conditions. There are many fingers to be pointed at who to blame for this problem, whether it be government’s fault or just personal responsibility. Most of the problems faced by the United States, such as obesity and diabetes, are the consequences of excessive unhealthy food intake. So many people in today’s society settle for the easiest and quickest way to get food, and usually end up at a fast food restaurant. Fast food is ridiculously cheap and easy to get. Little do they know, the food they are consuming is full of saturated fats, salts, and sugars which is very unhealthy. Fast food chains are known for their appeal to teenagers and kids, the risk of making children sick and unhealthy, and how it is easy to afford the cheap food they offer. Fast food chains such as McDonald’s and Burger King can be very appealing to children and teenagers in the 21st century. Both McDonald’s and Burger King both have a â€Å"mascot† so to speak; a clown and a king. In advertisements by both companies, both mascots are urging consumers to buy their product because that is what they do. If the ‘cool’ mascot does it, then nothing else matters. McDonald’s, especially, is the biggest appeal to children in all of fast food chains. They have Ronald McDonald, a big indoor play scape, and theyShow MoreRelatedThe Prevalence Of The Obesity Epidemic1677 Words   |  7 Pagesinsight on the raising obesity epidemic, caused mainly by the processed-food industry in North America. While the movie provides statistics on the obesity level, especially among children in the United States, it also indicates that the effects and influence of processed-food industry can be seen worldwide. 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The Importance of Macbeth in Modern Society Free Essays

The importance of continuing to read Macbeth in our modern society is to educate readers on valuable lessons in life. The significance of gaining power appropriately, the fact that there are consequences for every action and the importance of justice are a few of the many life lessons that are relevant in Shakespeare’s tragedy, Macbeth. Primarily, Macbeth, the main character, is influenced by his wife and subsequently has a great desire to be King of Scotland. We will write a custom essay sample on The Importance of Macbeth in Modern Society or any similar topic only for you Order Now Macbeth is presently the Thane of Glamis, as well as an army general of the current King, Duncan. When Duncan is in Inverness, he decides to stay at the home of the Macbeths. Lady Macbeth eagerly devises a plot to murder Duncan, thus ensuring her husband’s place on the throne of Scotland. Although Macbeth raises concerns about the regicide, he is eventually persuaded by his wife to follow her plan. â€Å" Stars, hide your fires! Let not light see my black and deep desires; the eye wink at the hand; yet let that be which the eye fears, when it is done, to see †(I. iv. 50-53). Macbeth knows that he must do wrong to achieve his goal, but plans to kill anyone in the way if he must. The method in which Macbeth chooses to use to gain power is neither civilized, nor humane. This extremity is perhaps hinting to the reader that Macbeth’s actions were wrong, as well as showing the extremes one may go through to get what they desire. The witches’ apparitions given to Macbeth and friend, Banquo, at the beginning of the plot play a monumental role in the thoughts of both men. Macbeth is told that he will be king, though Banquo’s sons will be kings as well. â€Å"As the weird woman promis’d; and, I fear, thou play’dst most foully for’t; yet it was said it should not stand in thy posterity, but that myself should be the root and father of many kings† (III.  I. 2-6). Banquo explains that he will father all kings to come, and that Macbeth will not be king for a long period of time. This apparition challenges the friendship between Macbeth and Banquo. Macbeth is not afraid to show his jealousy when saying, â€Å"Upon my head they plac’d a fruitless crown, and put a barren sceptre in my gripe, thence to be wrenc’d with an unlineal hand, no son of mine succeeding (III. I. 61-64). † Uneasy about the prophecy, Macbeth plans to kill Banquo and his son, Fleance, so that the premonition will not occur. Macbeth hires two men to murder Banquo and Fleance, but they only accomplished to kill Banquo, and Fleance is able to escape. The actions of Macbeth had consequences, as most actions do in present day. At the time of the murder, Macbeth does not think of the cost of his deeds. His self-interest and desire for the throne of Scotland drives him to finish the act. â€Å" Methought I heard a voice cry ‘Sleep no more! Macbeth does murder sleep† (II. ii. 35-36). Macbeth achieves power, but due to guilt, he cannot sleep and starts to drive himself mad. Macbeth was not the only person who was suffering from guilt. Lady Macbeth shows her agony when sleepwalking and talking to herself, â€Å"Out, damned spot! Out, I say! One; two: why, then ‘tis time to do’t. Hell is murky! Fie, my lord-fie! A soldier, and afeard? What need we fear who knows it, when none can call our power to account? † (IV. I. 34-37) Lady Macbeth frantically tries to wash her hands, but the reality is that there is nothing on them. It is evident that she is feeling guilty for her actions and because of it, she is going mad. Insanity and loss of sleep are only minute consequences, but play an immense role in the development of the characters throughout the play. Lastly, Macbeth achieves justice and what he deserved at the end of the play when Macduff beheads him. Macbeth and Macduff battle, sword for sword, but Macbeth boasts that he has no reason to fear Macduff, because of his belief in the witches’ prophesy. â€Å" Be bloody, bold, and resolute; laugh to scorn The power of man; for none of woman born Shall harm Macbeth† (IV. i. 86-87). This apparition informs Macbeth that he cannot be killed by any man â€Å"of woman born† (IV. i. 86-87). Macduff declares that he was â€Å"from his mother’s womb untimely ripp’d† which means that he was born by Caesarean section, and was therefore not â€Å"born† of a woman. The lesson to be learned is to not become a victim of peer pressure, and not to believe everything said. In conclusion, the importance of continuing to read Shakespeare’s Macbeth in our modern society is to educate readers about human behaviour on many valuable lessons in life. Whether the play teaches readers about gaining power appropriately, the fact that there are consequences for every action, or the importance of justice, it proves that they are important to learn. How to cite The Importance of Macbeth in Modern Society, Essay examples