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DNA

I INTRODUCTION  Deoxyribonucleic Acid (DNA), genetic material of all cellular organisms and most viruses. DNA carries the information needed to direct protein synthesis and replication. Protein synthesis is the production of the proteins needed by the cell or virus for its activities and development. Replication is the process by which DNA copies itself for each descendant cell or virus, passing on the information needed for protein synthesis. In most cellular organisms, DNA is organized on chromosomes located in the nucleus of the cell.

II STRUCTURE  
A molecule of DNA consists of two chains, strands composed of a large number of chemical compounds, called nucleotides, linked together to form a chain. These chains are arranged like a ladder that has been twisted into the shape of a winding staircase, called a double helix. Each nucleotide consists of three units: a sugar molecule called deoxyribose, a phosphate group, and one of four different nitrogen-containing compounds called bases. The four bases are adenine (abbreviated A), guanine (G), thymine (T), and cytosine (C). The deoxyribose molecule occupies the center position in the nucleotide, flanked by a phosphate group on one side and a base on the other. The phosphate group of each nucleotide is also linked to the deoxyribose of the adjacent nucleotide in the chain. These linked deoxyribose-phosphate subunits form the parallel side rails of the ladder. The bases face inward toward each other, forming the rungs of the ladder.


The nucleotides in one DNA strand have a specific association with the corresponding nucleotides in the other DNA strand. Because of the chemical affinity of the bases, nucleotides containing adenine are always paired with nucleotides containing thymine, and nucleotides containing cytosine are always paired with nucleotides containing guanine. The complementary bases are joined to each other by weak chemical bonds called hydrogen bonds.

In 1953 American biochemist James D. Watson and British biophysicist Francis Crick published the first description of the structure of DNA. Their model proved to be so important for the understanding of protein synthesis, DNA replication, and mutation that they were awarded the 1962 Nobel Prize for physiology or medicine for their work.

III PROTEIN SYNTHESIS  
DNA carries the instructions for the production of proteins. A protein is composed of smaller molecules called amino acids, and the structure and function of the protein is determined by the sequence of its amino acids. The sequence of amino acids, in turn, is determined by the sequence of nucleotide bases in the DNA. A sequence of three nucleotide bases, called a triplet, is the genetic code word, or codon, that specifies a particular amino acid. For instance, the triplet GAC (guanine, adenine, and cytosine) is the codon for the amino acid leucine, and the triplet CAG (cytosine, adenine, and guanine) is the codon for the amino acid valine. A protein consisting of 100 amino acids is thus encoded by a DNA segment consisting of 300 nucleotides. Of the two polynucleotide chains that form a DNA molecule, only one strand, called the sense strand, contains the information needed for the production of a given amino acid sequence. The other strand aids in replication.


Protein synthesis begins with the separation of a DNA molecule into two strands. In a process called transcription, a section of the sense strand acts as a template, or pattern, to produce a new strand called messenger RNA (mRNA). The mRNA leaves the cell nucleus and attaches to the ribosomes, specialized cellular structures that are the sites of protein synthesis. Amino acids are carried to the ribosomes by another type of RNA, called transfer RNA (tRNA). In a process called translation, the amino acids are linked together in a particular sequence, dictated by the mRNA, to form a protein.

A gene is a sequence of DNA nucleotides that specify the order of amino acids in a protein via an intermediary mRNA molecule. Substituting one DNA nucleotide with another containing a different base causes all descendant cells or viruses to have the altered nucleotide base sequence. As a result of the substitution, the sequence of amino acids in the resulting protein may also be changed. Such a change in a DNA molecule is called a mutation. Most mutations are the result of errors in the replication process. Exposure of a cell or virus to radiation or to certain chemicals increases the likelihood of mutations.

IV REPLICATION  
In most cellular organisms, replication of a DNA molecule takes place in the cell nucleus and occurs just before the cell divides. Replication begins with the separation of the two polynucleotide chains, each of which then acts as a template for the assembly of a new complementary chain. As the old chains separate, each nucleotide in the two chains attracts a complementary nucleotide that has been formed earlier by the cell. The nucleotides are joined to one another by hydrogen bonds to form the rungs of a new DNA molecule. As the complementary nucleotides are fitted into place, an enzyme called DNA polymerase links them together by bonding the phosphate group of one nucleotide to the sugar molecule of the adjacent nucleotide, forming the side rail of the new DNA molecule. This process continues until a new polynucleotide chain has been formed alongside the old one, forming a new double-helix molecule.

V TOOLS AND PROCEDURES  
Several tools and procedures facilitate the study and manipulation of DNA. Specialized enzymes, called restriction enzymes, found in bacteria act like molecular scissors to cut the phosphate backbones of DNA molecules at specific base sequences. Strands of DNA that have been cut with restriction enzymes are left with single-stranded tails that are called sticky ends, because they can easily realign with tails from certain other DNA fragments. Scientists take advantage of restriction enzymes and the sticky ends generated by these enzymes to carry out recombinant DNA technology, or genetic engineering. This technology involves removing a specific gene from one organism and inserting the gene into another organism.

Another tool for working with DNA is a procedure called polymerase chain reaction (PCR). This procedure uses the enzyme DNA polymerase to make copies of DNA strands in a process that mimics the way in which DNA replicates naturally within cells. Scientists use PCR to obtain vast numbers of copies of a given segment of DNA.

DNA fingerprinting, also called DNA typing, makes it possible to compare samples of DNA from various sources in a manner that is analogous to the comparison of fingerprints. In this procedure, scientists use restriction enzymes to cleave a sample of DNA into an assortment of fragments. Solutions containing these fragments are placed at the surface of a gel to which an electric current is applied. The electric current causes the DNA fragments to move through the gel. Because smaller fragments move more quickly than larger ones, this process, called electrophoresis, separates the fragments according to their size. The fragments are then marked with probes and exposed on X-ray film, where they form the DNA fingerprint—a pattern of characteristic black bars that is unique for each type of DNA.

A procedure called DNA sequencing makes it possible to determine the precise order, or sequence, of nucleotide bases within a fragment of DNA. Most versions of DNA sequencing use a technique called primer extension, developed by British molecular biologist Frederick Sanger. In primer extension, specific pieces of DNA are replicated and modified, so that each DNA segment ends in a fluorescent form of one of the four nucleotide bases. Modern DNA sequencers, pioneered by American molecular biologist Leroy Hood, incorporate both lasers and computers. The Human Genome Project, an international research collaboration, has been established to determine the sequence of all of the three billion nucleotide base pairs that make up the human genetic material.

An instrument called an atomic force microscope enables scientists to manipulate the three-dimensional structure of DNA molecules. This microscope involves laser beams that act like tweezers—attaching to the ends of a DNA molecule and pulling on them. By manipulating these laser beams, scientists can stretch, or uncoil, fragments of DNA. This work is helping reveal how DNA changes its three-dimensional shape as it interacts with enzymes.

VI APPLICATIONS  Research into DNA has had a significant impact on medicine. Through recombinant DNA technology, scientists can modify microorganisms so that they become so-called factories that produce large quantities of medically useful drugs. This technology is used to produce insulin, which is a drug used by diabetics, and interferon, which is used by some cancer patients. Studies of human DNA are revealing genes that are associated with specific diseases, such as cystic fibrosis and breast cancer. This information is helping physicians to diagnose various diseases, and it may lead to new treatments. For example, physicians are using a technology called chimeriplasty, which involves a synthetic molecule containing both DNA and RNA strands, in an effort to develop a treatment for a form of hemophilia.

Forensic science uses techniques developed in DNA research to identify individuals who have committed crimes. DNA from semen, skin, or blood taken from the crime scene can be compared with the DNA of a suspect, and the results can be used in court as evidence.

DNA has helped taxonomists determine evolutionary relationships among animals, plants, and other life forms. It is useful for this purpose, because closely related species have more similar DNA than do species that are distantly related. One surprising finding to emerge from DNA studies is that vultures of the Americas are more closely related to storks than to the vultures of Europe, Asia, or Africa.

Techniques of DNA manipulation are used in farming, in the form of genetic engineering and biotechnology. Strains of crop plants to which genes have been transferred may produce higher yields and may be more resistant to insects. Cattle have been similarly treated to increase milk and beef production, as have hogs, to yield more meat and less fat.

VII SOCIAL ISSUES  Despite the many benefits offered by DNA technology, some critics argue that its development should be monitored closely. One fear raised by such critics is that DNA fingerprinting could provide a means for employers to discriminate against members of various ethnic groups. Critics also fear that studies of people’s DNA could permit insurance companies to deny health insurance to those people at risk for developing certain diseases. The potential use of DNA technology to alter the genes of embryos is a particularly controversial issue.

The use of DNA technology in agriculture has also sparked controversy. Some people question the safety, desirability, and ecological impact of genetically altered crop plants. In addition, animal rights groups have protested against the genetic engineering of farm animals.

Despite these and other areas of disagreement, many people agree that DNA technology offers a mixture of benefits and potential hazards. Many experts also agree that an informed public can help assure that DNA technology is used wisely.

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