A clone is a cluster of individual entities or cells that are descended from one progenitor. One way to synthesize alkenes is by dehydration of alcohols, a process in which alcohols undergo E1 or E2 mechanisms to lose water and form a double bond. Note how the carbocation after the rearrangement is resonance stabilized by the oxygen. Draw a stepwise mechanism for the following reaction: 2c→4a+2b. The water molecule (which is a stronger base than the HSO4 - ion) then abstracts a proton from an adjacent carbon to form a double bond. This reaction is known as the Pinacol rearrangement.
For the production of vaccines like the hepatitis B vaccine. If the reaction is not sufficiently heated, the alcohols do not dehydrate to form alkenes, but react with one another to form ethers (e. g., the Williamson Ether Synthesis). Production of transgenic plants with improved qualities like insect and drought resistance and nutritional enrichment. The carbocation rearrangement would occur and determine the major and minor products as explained in the second part of this answer. Nitrogen fixation is carried out by cyanobacteria wherein desired genes can be used to enhance the productivity of crops and improvement of health. Assume no rearrangement for the first two product mechanisms. 3° alcohols: 25°– 80°C. Draw a stepwise mechanism for the following reaction: milady. They serve as a vehicle to carry a foreign DNA sequence into a given host cell. The Endonucleases cut within the DNA strand whereas the Exonucleases remove the nucleotides from the ends of the strands. However, the general idea behind each dehydration reaction is that the –OH group in the alcohol donates two electrons to H+ from the acid reagent, forming an alkyloxonium ion.
Application of Recombinant DNA Technology. It can be applied to the science of identifying and detecting a clone containing a particular gene which can be manipulated by growing in a controlled environment. They are two types, namely Endonucleases and Exonucleases. Recall that according to Zaitsev's Rule, the more substituted alkenes are formed preferentially because they are more stable than less substituted alkenes. Draw a stepwise mechanism for the following reaction: 2 h2 +. Gene cloning finds its applications in the agricultural field. Alcohols are amphoteric; they can act as both acid or base. This basic characteristic of alcohol is essential for its dehydration reaction with an acid to form alkenes. Also Refer- Gene Therapy. Secondary and tertiary alcohols dehydrate through the E1 mechanism. In every case the anionic leaving group is the conjugate base of a strong acid.
What is Recombinant DNA Technology? The E2 elimination of 3º-alcohols under relatively non-acidic conditions may be accomplished by treatment with phosphorous oxychloride (POCl3) in pyridine. The minor product being the same product as the one formed from the red arrows. 14.4: Dehydration Reactions of Alcohols. 2° alcohols: 100°– 140 °C. The effectively transformed cells/organisms carry forward the recombinant gene to the offspring. Scientists are able to generate multiple copies of a single fragment of DNA, a gene which can be used to create identical copies constituting a DNA clone. Listed below are the applications of gene cloning: - Gene Cloning plays an important role in the medicinal field. Similarly to the reaction above, secondary and tertiary –OH protonate to form alkyloxonium ions.
DNA technology is also used to detect the presence of HIV in a person. Thus the recombinant DNA has to be introduced into the host. Oxygen can donate two electrons to an electron-deficient proton. Notice in the mechanism below that the alkene formed depends on which proton is abstracted: the red arrows show formation of the more substituted 2-butene, while the blue arrows show formation of the less substituted 1-butene. So, basically, this process involves the introduction of a foreign piece of DNA structure into the genome which contains our gene of interest. The dehydration reaction of alcohols to generate alkene proceeds by heating the alcohols in the presence of a strong acid, such as sulfuric or phosphoric acid, at high temperatures.
The deprotonated acid (the base) then reacts with the hydrogen adjacent to the carbocation and form a double bond. Note: With the secondary carbocation adjacent a tertiary carbon center, a 1, 2 hydride shift (rearrangement) would occur to form a tertiary carbocation and vcompound below would be the major product. In the dehydration of this diol the resulting product is a ketone. Clinical diagnosis – ELISA is an example where the application of recombinant. The enzymes which include the restriction enzymes help to cut, the polymerases- help to synthesize and the ligases- help to bind. Additinally, trans alkenes are more stable than cis alkenes and are also the major product formed.
If there was a rearrangement, draw the expected major product. The vectors are made up of an origin of replication- This is a sequence of nucleotides from where the replication starts, a selectable marker – constitute genes which show resistance to certain antibiotics like ampicillin; and cloning sites – the sites recognized by the restriction enzymes where desired DNAs are inserted. The vectors – help in carrying and integrating the desired gene. Therapeutic protein production like insulin. The major product of this mechanism would be the more highly substituted alkene, or the product formed from the red arrows. These form a very important part of the tools of recombinant DNA technology as they are the ultimate vehicles that carry forward the desired gene into the host organism. Explain the roles of the following: (a) Restriction Enzymes. Also Refer: Genetically Modified Organisms (GMO). They are not part of the main cellular genome. This ion acts as a very good leaving group which leaves to form a carbocation. H2SO4 with heat since there are no concerns about C+ rearrangement.
For the example below, the trans diastereomer of the 2-butene product is most abundant. Medical ailments such as leukaemia and sickle cell anaemia can be treated with this principle. The lone pair of electrons on oxygen atom makes the –OH group weakly basic. The predominance of the non-Zaitsev product (less substituted double bond) is presumed due to steric hindrance of the methylene group hydrogen atoms, which interferes with the approach of base at that site. The more substituted alkene is favored, as more substituted alkenes are relatively lower in energy. Plasmids are circular DNA molecules that are introduced from bacteria. Ligation of DNA Molecules. This gene which is introduced is the recombinant gene and the technique is called the recombinant DNA technology. Plasmids and bacteriophages are the most common vectors in recombinant DNA technology that are used as they have a very high copy number. As mentioned in Tools of recombinant DNA technology, there are various ways in which this can be achieved. There are a number of ways in which these recombinant DNAs are inserted into the host, namely – microinjection, biolistics or gene gun, alternate cooling and heating, use of calcium ions, etc.
The desired genes and the vectors are cut by the same restriction enzymes to obtain the complementary sticky notes, thus making the work of the ligases easy to bind the desired gene to the vector. Starting with cyclohexanol, describe how you would prepare cyclohexene. Insertion of Recombinant DNA Into Host. B) Plasmid is an extra-chromosomal DNA molecule in bacteria that is capable of replicating, independent of chromosomal DNA. Primary alcohols undergo bimolecular elimination (E2 mechanism) while secondary and tertiary alcohols undergo unimolecular elimination (E1 mechanism). Gene therapy in diseases like cancer, SCID etc. It is a process to amplify a single copy of DNA into thousands to millions of copies once the proper gene of interest has been cut using restriction enzymes. Frequently Asked Questions. It carries genes, which provide the host cell with beneficial properties such as mating ability, and drug resistance.
Dehydration of Alcohols to Yield Alkenes. Host organism – into which the recombinant DNA is introduced. Discuss the applications of recombination from the point of view of genetic engineering. Also Read: Bioinformatics.
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