4 Selective BreedingHumans use selective breeding, which takes advantage of naturally occurring genetic variation in plants, animals, and other organisms, to pass desired traits on to the next generation of organisms
5 Selective BreedingNearly all domestic animals -- including horses, cats, and farm animals – and most crop plants have been produced by selective breeding
6 HybridizationLouis Burbank was the greatest selective breeder of all time. He developed the disease-resistant potato and more than 800 varieties of plants.
7 Louis Burbank used the technique of hybridization and bred dissimilar individuals to combine the best traits of both parents.The hybrids produced by these crosses were hardier than their parents
8 InbreedingTo maintain the desired characteristics of a line of organisms, breeders often use the technique of inbreeding.Inbreeding is the continued breeding of individuals with similar characteristics
9 Increasing VariationIn order for selective breeding to be successful, there must be a lot of genetic variation in the populationBreeders increase the genetic variation in a population by inducing mutations, which are the ultimate source of genetic variability
10 Increasing VariationBreeders increase the mutation rate by using radiation and chemicals
11 Producing New Kinds of Bacteria and Plants Using radiation or chemicals, scientists have been able to develop hundreds of useful bacteria strainsDrugs that prevent chromosomal separation during meiosis have been used to produce plants that have many sets of chromosomes.The hybrid plants are polyploid.
12 What is genetic engineering? In 1973, Mr. Cohen and Mr. Boyer conducted an experiment on the DNA of an American frog.
13 What is genetic engineering? They found and isolated the gene that codes for ribosomal RNA (rRNA) in the DNA of the frog. They removed that gene from the frog and inserted it into some E. Coli Bacteria.
14 What Happened?During transcription, the bacteria then produced the frog RNA!Genetic Engineering: the process of manipulating (moving) genes for practical purposes (useful)Recombinant DNA: DNA made from 2 or more organisms that are different.
15 The Basic Steps of Genetic Engineering Cutting the DNA:Restriction Enzymes: bacterial enzymes that recognize and bind to specific short sequences of DNA, and then cut the DNA between specific nucleotides within the sequences.Vector: agent used to carry the gene of interest – usually plasmidsPlasmid: the circular DNA molecules that replicate
16 The Basic Steps to Genetic Engineering Making Recombinant DNADNA fragments of interest (that have already been cut) are combined with the vector.DNA ligase – the enzyme bonds the 2 ends of the fragments to the vectors.CloningGene cloning: the process of making many copies of a geneBacteria reproduce by binary fission
17 The Basic Steps to Genetic Engineering ScreeningCells that have received the gene of interest are separated out.Those cells then continue to produce the protein coded for by the gene
18 Cutting DNA & Making Recombinant DNA How Restriction enzymes work:The Enzymes recognize specific sequences on Human and Bacterial PlasmidsThe Enzymes cut the strands.The cut produces DNA fragments with short strands on each end that are complementary to each other“Sticky Ends”Both the human DNA and the Plasmid “Open Up” with the same sticky ends remainingThey Bind Together
20 Recognition sequences DNA sequenceRestriction enzyme EcoRI cuts the DNA into fragments.Sticky end
21 Confirmation of a Cloned Gene One method used identify a specific gene is called a Southern BlotSteps:Cut DNA from bacteria with restriction enzymes.DNA fragments are separated by a gel soaked in a chemical solution.Gel electrophoresis – uses an electric field within a gel to separate molecules by their size
22 Confirmation of a Cloned Gene Negatively charged DNA is put into these wells.They are attracted to the positive pole from the electric field.The Smallest DNA fragments move the fastest
23 Gel Electrophoresis DNA plus restriction enzyme Power source Longer fragmentsShorter fragmentsMixture of DNA fragmentsGel
24 Confirmation of a Cloned Gene The DNA separated is then transferred to a filter paper (blotted) and a probe solution is added.Probes: radioactive RNA or single-stranded DNA pieces that are complementary to the gene of interestOnly DNA fragments complementary to the probe will form and bind bands
25 Confirmation of Cloned Genes Why do this?Bacterial colonies can be used to produce large quantities of the protein (used to study or make drugs)
26 Confirmation of Cloned Genes When a bacteria or other cell takes in a foreign piece of DNA such as a plasmid, the process is called transformationIf transformation is successful, the recombinant DNA is integrated into one of the chromosomes of the cell.
27 Improving CropsGenetic engineers can add favorable characteristics to a plantPlants become resistant to insects (no longer need pesticides); resistant to weed killer (so crops won’t die, but weeds will); improved nutrition – rice + corn
28 Plant TransformationInside plant cell, Agrobacterium inserts part of its DNA into host cell chromosomeAgrobacterium tumefaciensGene to be transferredCellular DNARecombinant plasmidPlant cell coloniesComplete plant is generated from transformed cellTransformed bacteria introduce plasmids into plant cells
29 Animal FarmingGrowth hormones is given to cows to produce more milkHuman genes are added to farm animals in order to have human proteins in their milkThe Human proteins are extracted from milk and sold to pharmacy companies.Useful for complex proteins that can’t be made in bacteria
30 Creating HGH Human Cell Bacterial Cell Plasmid Gene for human growth hormoneRecombinant DNAGene for human growth hormoneHuman CellDNA recombinationSticky endsDNA insertionBacterial CellBacterial chromosomeBacterial cell for containing gene for human growth hormonePlasmid
31 Animal FarmingTransgenic animals: Animals that have foreign DNA in their cellsCloning of animals is another way to make large quantities of a certain protein.
32 Animal Farming Transgenic animals: How it works: an intact nucleus from an embryonic cell (whose DNA has recombined with a human gene) is placed into an egg whose nucleus has been removed.The “new” egg is then placed into the uterus of an animal.
33 Cloning AnimalsA clone is a member of a population of genetically identical cells produced from a single cellHow it works: an intact nucleus from a cell is removed
34 Cloning AnimalsThe nucleus is fused with a egg cell (whose nucleus has been removed) taken from another adultThe fused cell begins to divide and the embryo is placed in the uterus of a foster mother.The “new” egg is then develops normally.
36 Cloning Embryo A donor cell is taken from a sheep’s udder. Donor NucleusThese two cells are fused using an electric shock.Fused CellEgg CellThe nucleus of the egg cell is removed.The fused cell begins dividing normally.An egg cell is taken from an adult female sheep.Cloned LambThe embryo is placed in the uterus of a foster mother.EmbryoFoster MotherThe embryo develops normally into a lamb—Dolly
37 Genetically Engineered Drugs and Vaccines Today, many pharmaceutical companies around the world produce important proteins using genetic engineering.Vaccine: a solution containing all or part of a harmless version of a pathogen; used to prevent viral diseases (don’t respond to drugs)Many vaccines are made using genetic engineering
38 DNA FingerprintingDNA fingerprinting:a pattern of dark bands on photographic filman individuals’ DNA restriction fragments are separated by gel electrophoresis, probed, and exposed to X-ray film.
39 DNA fingerprints can be used to DNA FingerprintingDNA fingerprints can be used toestablish paternityidentify genetic disordersforensics (scientific study of cause of injury or death in criminal activity)
40 Three paternity cases tested with DNA purified from blood samples Three paternity cases tested with DNA purified from blood samples. 1: mother; 2: child; 3: alleged father; 4: child + alleged father. M: markers. 3 µg of DNA was loaded per lane. Outcome: A: alleged father excluded; B & C: alleged father confirmed.(Data kindly provided by J. James, Gene Proof Technologies, Nashville, TN, USA.)