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Biotechnology. Cloning. Production of genetically identical copies of DNA, cells or organisms Asexual reproduction Gene therapy – genes used to modify a human Transgenic organisms. Steps of Genetic Engineering. Cohen and Boyer 1973 1 st successful GE experiment frog rRNA into E. coli
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Cloning • Production of genetically identical copies of DNA, cells or organisms • Asexual reproduction • Gene therapy – genes used to modify a human • Transgenic organisms
Steps of Genetic Engineering • Cohen and Boyer • 1973 1st successful GE experiment • frog rRNA into E. coli • Steps: • Cleave DNA • Recombinant DNA • Clone DNA • Screen DNA
Recombinant DNA • rDNA – contains DNA from 2 or more sources • Vector (plasmid or virus) used to transfer gene of interest to host cell • Restrictive enzymes and DNA ligase are used to cut and seal the ends of the gene of interest.
Bacteria cells do not have introns. • For successful genetic engineering exp., the gene must not introns. • RNA transcriptase can make DNA from mRNA = cDNA (DNA without introns)
Polymerase Chain Reaction • Creates copies of small segments of DNA • Uses DNA polymerase and nucleotides • Only needs small sample • Can be used to study phylogeny of human populations
DNA fingerprinting • Usually follows PCR • Add restrictive enzymes to sample of DNA to make many different sized fragments • Gel electrophoresis is used to separate the fragments • If two banding patterns match, likelihood the DNA came from same person • Used in convicting criminals, detecting cancer, viral infections or genetic disorders, identifying remains of bodies
Biotechnology products • Genetically modified in order to produce a product desired by humans!!!
Transgenic Bacteria • Insulin, clottng factor VIII, human growth hormone, t-PA, hepatitis B vaccine • Frost resistant strawberries, root protection from insects • Enhancing ability to degrade substances • Clean up oil, remove sulfur from coal, clean up toxic wastes • Self destruct when job is done
Transgenic plants • Foreign genes are placed in immature plant embryos or protoplasts (plant cells with cell wall removed) • Pomato (potatoes and tomatoes) • Cotton, corn and potatoes resistant to pests • Soybeans – herbicide resistant • Producing human proteins (hormones, clotting factors and antibodies)
Transgenic Animals • Insert genes into the eggs of animals • Can be done by hand or by vortex mixing • bGH – bovine growth hormone – makes larger fish, cows, pigs, rabbits, sheep • Gene pharming – use of transgenic farm animals to make pharaceuticals • Genes that code for medical proteins are put into animal DNA and appear in milk
Genomics • Study of genomes – genes • Human genome project – sequenced the base pairs in the DNA of the human chromosome. • HapMap Project • Goal – catalog common sequence differences that occur in humans, find risk for specific illnesses • Haplotypes – patterns of sequence differences
Genetic profile • Identify the complete genotype of an individual, including mutations • DNA chip produced by a quick swab • Can help with the way we live our lives, preventive measures
Proteomics • Study of the structure, function and interaction of cellular proteins • Human proteome – translation of genes into proteins
Bioinformatics • Application of computer technologies to the study of the genome • Goal – find cause/effect relationship between genetic profiles and genetic disorders caused by multifactorial genes.
Gene Therapy • Insertion of genetic material into human cells for the treatment of a disorder. • Ex Vivo Gene Therapy • Cells are removed, manipulated, then replaced • Used in treatment of SCID and hypercholesterolemia • In Vivo Gene Therapy • Directly insert healthy genes to replace faulty ones • Cystic fibrosis, VEGF, gene p53 and cancer