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Cloning of a Gene Biotechnology Recombinant DNA Technology • Cloning is the production of identical copies through asexual means. • Cloning occurs naturally in new plant shoots, bacterial colonies, and identical human twins. • Gene cloning is the engineering and thus production of many identical copies of a genes Cloning a human gene • Recombinant DNA contains DNA from two or more different sources. – Recombinant DNA can be the recombination of parents DNA to form a hybrid offspring (you and I are an example of a hybrid) • A plasmid can be used as a to insert foreign DNA into a cell – A plasmid is a small ring of DNA transmitted between bacteria that live close to one another Cloning of a Gene • Restriction enzymes “cut” the recombinant DNA and the source DNA at a specific sequence, leaving “sticky” ends, that allow a portion of source DNA to be inserted into the vector DNA. • Bacterial cells take up recombinant plasmids and clone the new DNA 1 The Polymerase Chain Reaction The Polymerase Chain Reaction • The polymerase chain reaction (PCR) produces many copies of a single gene or piece of DNA. • PCR requires DNA polymerase and a supply of nucleotides for the new DNA strands. • PCR is a chain reaction because the targeted DNA is repeatedly replicated as long as the process continues. Analyzing DNA Segments • The entire genome of an individual can be cut by restriction enzymes to yield variable fragment lengths. • Gel electrophoresis separates fragment lengths and the use of probes results in a pattern unique to the individual, sometimes called a DNA fingerprint. Cloning of a Gene – A DNA fingerprint resembles that of one’s parents because it is inherited. • It’s possible to identify a person who has committed a crime, identify a cancer gene, or tell who is related to whom by performing a DNA fingerprint. • Since PCR can amplify the smallest amount of DNA, a single sperm, or one cell on a toothbrush, provides enough DNA to be identified by comparison with sample DNA. Biotechnology Products Transgenic Bacteria • Today, bacteria, plants, and animals are genetically engineered to make biotechnology products. • Organisms that have had a foreign gene inserted into them are called transgenic organisms. • There are now transgenic bacteria, transgenic plants, and transgenic animals. • Recombinant DNA technology is used to produced transgenic bacteria, which are grown in huge vats called bioreactors. • Transgenic bacteria are used to produce: insulin, human growth hormone, tissue plasminogen activator, and hepatitis B vaccine. • They also add insecticidal toxins to plants, reduce frost damage on plants, degrade wastes, produce chemicals, and help mine metals. 2 Transgenic Plants Transgenic Animals • Foreign genes are added to protoplasts using an electric current. • Foreign genes in cotton, corn, and potatoes have given them pest resistance; soybeans are made resistant to herbicide for no-till farming. • Transgenic plants produce human hormones, clotting factors, and antibodies in their seeds; one weed has even been engineered to produce plastic granules. • Foreign genes can be inserted into animal eggs. • Gene pharming uses transgenic farm animals to produce pharmaceuticals in their milk. • Animals may produce drugs for the treatment of cystic fibrosis, cancer, and blood diseases. Transgenic animals Cloning Transgenic Animals • Cloning of mammals was once considered impossible, but has now been accomplished with sheep, calves and goats. • A 2n nucleus from a bioengineered animal is inserted into enucleated eggs from a donor. • A surrogate mother gives birth to the cloned animals. Animal Organs as Biotechnology Products • Scientists are genetically engineering pigs to serve as organ donors for humans who need transplants. • Transplants of organs across species is called xenotransplantation. • Researchers are trying to make organs less antigenic to humans. The Human Genome Project The Human Genome Project has two goals: (1) to construct a map that shows the sequence of bases on all the human chromosomes and (2) to construct a map that shows the sequence of genes along all the human chromosomes. 3 The Base Sequence Map Genetic map of chromosome 17 • The first goal has been completed and researchers know the sequence of three billion base pairs after 15 years of research. • The two agencies that completed the task are The International Human Genome Sequencing Consortium and Celera Genomics, a private company. The Base Sequence Map • A gene map could help discover mutant genes and lead to the development of medicines to treat these disorders. • Germline therapy, that is done before a child is born, may eventually be possible. • Many ethical questions surround how human genome maps should be used. Gene Therapy • Gene therapy is the insertion of genetic material into human cells for the treatment of a disorder. • A patient would be given healthy genes to make up for any faulty genes. • Many researchers are trying to cure cancer by inserting genes to make healthy cells tolerant of chemotherapy or use a gene (p53) to bring about the death of cancer cells. 4