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APPLIED GENETICS SELECTIVE BREEDING • Breeders chose organisms with desirable traits to be the parents of the next generation. • Requires time, patience and many generations before desired trait becomes common in a population. • Examples: cows, chickens, largest grain heads, largest fruits. I AM THE GREATEST!! Breeding for desirable traits I’M A HANDSOM E SPECIMEN I’M SO PRETTY INBREEDING • Mating between closely related individuals • Ensures offspring are homozygous for most traits. • However, also brings out harmful recessive traits since closely related organisms are more likely to carry recessive alleles for traits. • Pure breeds: dogs, horses • A breed/cultivar: selected group of organisms within species that has been breed for a particular characteristic. Recessive traits appear! • The dachshund breed of dogs was developed to dig out rodents however it’s long slim body also makes it prone to slipped or ruptured disks in its vertebrae. • The large dogs such as St. Bernard’s have a problem with their hip bones developing arthritis as the dog gets old because of their size. MY BACK HURTS! HYBRIDIZATION • Hybrids are produced by crossing two unlike organisms. • Corn - bred for high yield (domestic varieties) and disease resistance (wild varieties - maize) • Mule - female horse x male donkey – bred for size and strength – STERILE WE CAN’T HAVE BABIES? DETERMINING GENOTYPE • Test cross is used to determine parental genotype. • Cross the parent with unknown genotype with an organism that is homozygous recessive for the trait. • Observe the offspring. • Numerous test crosses required to be sure you findings are correct. RECOMBINANT DNA TECHNOLOGY GENETIC ENGINEERING TOOLS Electrophoresis Chamber DNA molecule PLASMID GENETIC ENGINEERING • Or Recombinant DNA Technology • Fast, more reliable method for increasing frequency of specific allele in a population. • A small fragment of DNA is cut from one organism and inserted into another host organism of a different species • Recombinant DNA forms when DNA fragments from different species is combined. • Resulting transgenic organisms contain functional recombinant DNA 3 STEPS TO MAKE A TRANSGENIC ORGANISM 1. ISOLATION OF FOREIGN DNA: a specific restriction enzyme is used to cut the required fragment of DNA 2. ATTACH DNA FRAGMENT TO A VECTOR: for transportation into the host. Vectors maybe plasmids, viruses, tiny metal bullets, micropipettes. (Gene splicing in plasmids.) 3. TRANSFER OF VECTOR INTO HOST ORGANISM - creating a transgenic organism. A transgenic organism reproduces a copy of the recombinant DNA, when it divides, and can use that gene as it was used by the original organism. APPLICATIONS OF DNA TECHNOLOGY 3 MAIN AREAS OF USE 1. INDUSTRY 2. MEDICINE 3. AGRICULTURE TRANSGENIC BACTERIA INDUSTRIAL USES • • Breaking down pollutants to harmless products Degradation of spilled oil, extraction of minerals from ores, bioremediation. MEDICINAL USES • • Production of hormones: growth hormone, insulin Producing phenylalanine for artificial sweeteners AGRICULTURAL USES • • Reduction of frost damage to plants Increased rate of nitrogen fixation in the soil TRANSGENIC PLANTS • Plants are more difficult to genetically engineer than bacteria because 1. No plasmids 2. Thick cell wall 3. Mechanical vectors commonly used • Engineered plants may: 1. Resist herbicides 2. Produce internal pesticides 3. Increase protein production TRANSGENIC ANIMALS • • • Used for studying diseases and roles played by specific genes with a view to curing some human diseases Commonly used animals mice, roundworms and fruit flies Examples: 1. Goats engineered to produce high levels of human protein to dissolve blood clots. 2. Cows engineered to produce higher milk yields CLONING • Cloning is a type of genetic engineering in which an exact duplicate of an organism is created from a single body cell. • This is done in trees to produce many organisms from a single organism in order to reforest certain areas • It has only been the last few years that it has been possible in mammals as well. This type of genetic engineering is artificial and done in a laboratory. Dolly THE HUMAN GENOME HUMAN GENOME PROJECT • Organized in 1990, completed in 2003 • Worldwide collaborative project • Objective was to completely map and sequence the human genome (3 billion base pairs in 20,000-25,000 genes on 46 chromosomes) SEQUENCING DNA • Allows scientists to identify and study specific genes and fragments of DNA that regulated the transcription of the genes. • Millions of copies of the DNA fragment are cloned and fragmented further for analysis. • Analyzed using GEL ELECTROPHORESIS, produces a pattern of dyed bands on a gel. LINKAGE MAPS • Show the location of genes on a chromosome • Historically produced by analyzing crossed-over traits in families: inefficient due to few offspring and generation time. • Genes are now mapped using PCR polymerase chain reaction. • Millions of copies of DNA fragments are cloned in hours from sperm cells of one person and analyzed for crossing over. • Machines are used for the analysis APPLICATIONS OF THE HUMAN GENOME PROJECT DIAGNOSIS OF GENETIC DISORDERS • DNA from people suffering from a disorder are analyzed for common patterns • Fetal cells are collected • Fetal cells are grown on a cell culture • Fetal cell DNA analyzed for pattern associated with disorder • A gene at site 4p16.3 was found which had a variable trinucleotide repeat (CAG). This gene is directly related to Huntington's Disease and the production of the protein "huntingdin". The role of this protein is not yet know, much to the chagrin of researchers. GENE THERAPY • Insertion of normal genes into human cells to correct disorder • CF: see picture • SCIDS: Missing an enzyme due to faulty gene. Cells are removed, enzyme producing gene incorporated and return to patient. (Genes do not remain active long term) DNA FINGERPRINTING • Shows the DNA sequences of an individual • DNA samples from hair blood skin semen are copied using PCR. • DNA cut into fragments of different lengths using restriction enzymes • Fragments are separated using gel electrophoresis • DNA fingerprints produced are matched • NO two individuals have the same DNA fingerprints