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Dosya Açıklaması : DNA is a polymer. The monomer units of DNA are nucleotides, and the polymer is known as a "polynucleotide." Each nucleotide consists of a 5-carbon sugar (deoxyribose), a nitrogen containing base attached to the sugar, and a phosphate group. There are four different types of nucleotides found in DNA, differing only in the nitrogenous base. The four nucleotides are given one letter abbreviations as shorthand for the four bases. • A is for adenine • G is for guanine • C is for cytosine • T is for thymine Adenine and guanine are purines. Purines are the larger of the two types of bases found in DNA. Cytosine and thymine are pyrimidines. Like purines, all pyrimidine ring atoms lie in the same plane. The deoxyribose sugar of the DNA backbone has 5 carbons and 3 oxygens. The hydroxyl groups on the carbons link to the phosphate groups to form the DNA backbone. Deoxyribose lacks an hydroxyl group when compared to ribose, the sugar component of RNA. DNA is a normally double stranded macromolecule. Two polynucleotide chains, held together by weak thermodynamic forces, form a DNA molecule. Features of the DNA Double Helix are: • Two DNA strands form a helical spiral, winding around a helix axis in a right-handed spiral • The two polynucleotide chains run in opposite directions • The sugar-phosphate backbones of the two DNA strands wind around the helix axis like the railing of a sprial staircase The bases of the individual nucleotides are on the inside of the helix, stacked on top of each other like the steps of a spiral staircase. Introduction Although scientists as far back in history as Aristotle recognized that the features of one generation are passed on to the next (...like begets like...) it was not until the 1860s that the fundamental principles of genetic inheritance were described by Gregor Mendel. His theories were, however, widely disregarded by scientists of the time. In the last quarter of the 19th century, however, microscopists and cytologists, interested in the process of cell division, developed both the equipment and the methods needed to visualize chromosomes and their division in the processes of mitosis and of meiosis. As the 20th century began many scientists noticed similarities in the theoretical behavior of Mendels particles, and the visible behavior of the newly discovered chromosomes. It wasnt long before most scientists were convinced that the hereditary material responsible for giving living things their characteristic traits, and chromosomes must be one in the same. Yet, questions still remained. Chemical analysis of chromosomes showed them to be composed of both protein and DNA. Which substance carried the hereditary information? For many years most scientists favored the hypothesis that protein was the responsible molecule because of its comparative complexity when compared with DNA. After all, DNA is composed of a mere 4 subunits while protein is composed of 20, and DNA molecules are linear while proteins range from linear to multiply branched to globular. It appeared clear that the relatively simple structure of a DNA molecule could not carry all of the genetic information needed to account for the richly varied life in the world around us! In 1951, the then 23-year old biologist James Watson traveled from the United States to work with Francis Crick, an English physicist at the University of Cambridge. Crick was already using the process of X-ray crystallography to study the structure of protein molecules. Together, Watson and Crick used X-ray crystallography data, produced by Rosalind Franklin and Maurice Wilkins at Kings College in London, to decipher DNAs structure. Working with nucleotide models made of wire, Watson and Crick attempted to put together the puzzle of DNA structure in such a way that their model would account for the variety of facts that they knew described the molecule. Once satisfied with their model, they published their hypothesis, entitled "Molecular Structure of Nucleic Acids: A Structure for Deoxyribose Nucleic Acid" in the British journal Nature. It is interesting to note that this paper has been cited over 800 times since its first appearance! Here are their words: "...This (DNA) structure has two helical chains each coiled round the same axis...Both chains follow right handed helices...the two chains run in opposite directions. ..The bases are on the inside of the helix and the phosphates on the outside..." "The novel feature of the structure is the manner in which the two chains are held together by the purine and pyrimidine bases... The (bases) are joined together in pairs, a single base from one chain being hydrogen-bonded to a single base from the other chain, so that the two lie side by side...One of the pair must be a purine and the other a pyrimidine for bonding to occur. ...Only specific pairs of bases can bond together. These pairs are: adenine (purine) with thymine (pyrimidine), and guanine (purine) with cytosine (pyrimidine)." "...in other words, if an adenine forms one member of a pair, on either chain, then on these assumptions the other member must be thymine; similarly for guanine and cytosine. The sequence of bases on a single chain does not appear to be restricted in any way. However, if only specific pairs of bases can be formed, it follows that if the sequence of bases on one chain is given, then the sequence on the other chain is automatically determined." "...It has not escaped our notice that the specific pairing we have postulated immediately suggests a possible copying mechanism for the genetic material." And with these words, the way was made clear for tremendous strides in our understanding of the structure of DNA and, as a result our ability to work with and manipulate the information-rich DNA molecule. 1.0. Historical Past of Knowledge about DNA: 1.1 From 1870s to Watson-Crick: The processes of mitosis and meiosis were discovered in the 1870s and 1890s. It was observed that, as cells divided, chromosomes moved around in a cell, and people began to wonder what their function was. It was determined that chromosomes were made of protein and a chemical called DNA, about which people knew almost nothing. People began to suspect that chromosomes had something to do with genetics, but couldnt explain what/how. When enough evidence was accumulated to confirm that chromosomes did, indeed, have something to do with genetics, most people thought that in some way the protein in the chromosomes served as the genetic material. People knew that DNA was also in the chromosomes, but because its structure was unknown and people didnt know much about it, few people thought it was the genetic material. In 1928, Frederick Griffith performed an experiment using pneumonia bacteria and mice. This was one of the first experiments that hinted that DNA was the genetic code material. Follow the links to study his experiment. He used two strains of Streptococcus pneumoniae: a "smooth" strain which has a polysaccharide coating around it that makes it look smooth when viewed with a microscope, and a "rough" strain which doesnt have the coating, thus looks rough under the microscope. When he injected live S strain into mice, the mice contracted pneumonia and died. When he injected live R strain, a strain which typically does not cause illness, into mice, as predicted, they did not get sick, but lived. Thinking that perhaps the coating on the bacteria somehow caused the illness, Griffith then used heat to kill some of the S strain bacteria and injected that into mice. This failed to infect/kill the mice, indicating that the polysaccharide coating was not what caused the disease, but rather, something within the living cell. Since Griffith had used heat to kill the bacteria and heat denatures protein, he next hypothesized that perhaps some protein within the living cells, that was denatured by the heat, caused the disease. He then injected another group of mice with a mixture of heat-killed S and live R, and the mice died! When he did a necropsy on the dead mice, he isolated live S strain bacteria from the corpses. Griffith concluded that therefore, the live R strain bacteria must have absorbed genetic material from the dead S strain bacteria, and since heat denatures protein, the protein in the bacterial chromosomes was not the genetic material. This evidence pointed to DNA as being the genetic material. Transformation is the process whereby one strain of a bacterium absorbs genetic material from another strain of bacteria and "turns into" the type of bacterium whose genetic material it absorbed. Because DNA was so poorly understood, scientists remained skeptical up through the 1940s In 1952, Alfred Hershey and Martha Chase did an experiment which is so significant, it has been named the "Hershey-Chase Experiment". They used a bacterium named Escherichia coli, or E. coli for short, named after a scientist whose last name was Escher. They also used a virus called T2 that infects E. coli. T2 is a bacteriophage. Isolated T2, like other viruses, is just a crystal of DNA and protein, so it must live inside E. coli in order to make more virus like itself. When the new T2 viruses are ready to leave the host E. coli cell (and go infect others), they burst the E. coli cell open, killing it (hence the name "bacteriophage"). Hershey and Chase were seeking an answer to the question, "Is it the viral DNA or viral protein coat (capsid) that is the viral genetic code material which gets injected into the E. coli?" Their results indicated that the viral...