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BIOTECHNOLOGY-Genetic Engineering, Restriction Digestion, and Electrophoresis What is Biotechnology? Any technique that uses living organisms or substances from those organisms to make or modify __________________________________________________ improve ___________________________________________ to develop ____________________________for specific uses Genetic engineering is taking one or more ____________________from one organism and 1. Transferring them to _____________________________________________(most common) 2. Putting them back into the original organism in _____________________________________ i. Transgenic Organisms: Bt Corn (____________________________________) ii. Recombinant DNA (rDNA): ________________________________________ What Are the Benefits of Biotechnology? __________________, Human, Veterinary, Environment, Agriculture, __________________________, Industry and Manufacturing rInsulin The first commercial product made by genetic engineering (________). Insulin is the __________________ used to control their ________________ levels in ___________________________ First sold in ___________, (S. San Francisco) Biotech Tools Restriction Enzyme: ___________________________________________ Ligase: ______________________________________________________ ___________________________(Re-establishes the phosphodiester bond) Gene of Interest: _____________________________________________ Recipient Genome: ___________________________________________ Plasmid: __________________________ that bacteria have in addition to ___________________________________________________ EXAMPLE Restriction Enzyme Natural function of restriction enzymes: 1. ___________________________________________________________________________ 2. Digest foreign (e.g. viral) DNA. 3. ___________________________________________________________________________ A closer look…. EcoR1 5’….ACTGTACGAATTCGCTA….3’ 3’….TGACATGCTTAAGCGAT….5’ The DNA Ligase ________________________________ Let’s try it by creating a “Paper Plasmid” HOW ENGINEERING A PLASMID REALLY DONE? Steps for genetically engineering INSULIN 1. Take a _____________________________ and remove the nucleus 2. Extract the ________________________(genome) from the nucleus 3. Locate the _____________________________________________ 4. Cut out the Insulin gene with a _____________________________ 5. Take a _______________________________ and cut open with the ____________________________________________ 6. Place the _________________ into the bacterial _________________; you now _________________________________________________. 7. Place the ____________________________ back into an bacterial cell (transformation) 8. The Bacteria will now produce ____________________ as a by-product. WHAT ELSE CAN WE DO WITH DNA Gel electrophoresis- Cuts DNA at repetitive sections that are unique=called Restriction Fragment Length Polymorphisms (RFLPs) -We can use the fragments made by the _____________________________________ to make a ___________________________________________ -Looking for about ______ variations -DNA is ___________________________ charged (due to ________________________________) -When we pump electrical charge into the buffer, it will make the DNA move toward the positive charge______________________________________________ Imagine you are a DNA molecule -Smaller DNA fragments travel ____________________________________________ -If you were inside an agarose gel, your environment would resemble a _________________________. -The smaller the DNA fragment, the ____________________it is to get through the web. -This creates a DNA Fingerprint http://www.dnalc.org/ddnalc/resources/electrophoresis.html Sample Gel Who Committed the Crime? How do you know? LET’S TRY IT Crime Scene Activity (Tape= _________________________, Scissors= ________________________) Lab: Crime Scene Analysis Restriction digestion with BamH1 (________________________) -Use Blue Pen Restriction digestion with EcoR1 (_________________________) -Use Red Pen Plot fragments on _______________________________________________ (based on fragment sizes) Exclude suspects Answer questions on mock gel WS Answer wrap questions Find news article on DNA fingerprinting and summarize article What is Cloning? Clones are organisms that are ________________________________. Every single bit of their DNA is identical. Clones can happen naturally—______________________________________ are just one of many examples. Or they can be made in the lab by Somatic Cell Nuclear Transfer Cloning vs. Cloning a gene When scientists clone an organism, they are making an exact genetic copy of the ________________________ Cloning a gene usually involves copying the __________________________ of that gene into a smaller, more easily manipulated piece of DNA, such as a ________________________. This process makes it easier to study the function of the individual ___________________________. Somatic Cell Nuclear Transfer (SCNT) aka: __________________________ 1952 – _______________________________ 1996 – The ____________________________ cloned from adult cells was ___________________________. 2001 – The first human embryo cloned, but on divided to ______________________, providing evidence that human cloning is ___________________________________. Dolly: ________________tries= ___________Sheep On 14 February 2003, Dolly was ____________________ because she had a progressive lung disease and severe arthritis. Normal Life expectancy: _____________________ Let’s Click and Clone http://learn.genetics.utah.edu/content/tech/cloning/clickandclone/ EVIDENCE FOR EVOLUTION: 1. Fossils show change over time scientists can ___________________ & use them to support the _______________________________ ___________________________revel whether species are related. Anatomy of ____________________ also shows relatedness 2. Homologous Structures ________________________________________ because they share a common ancestor: These structures “ ________________________” Ex: human arm, dog front limb, horse leg, whale 3. Analogous structures Distantly related species have _______________________________ but are ________________ in structure _________________________ These structures “ ________________________” Ex: wing of butterfly & bird 4. Vestigial structures Structures ________________ & often ____________________ ___________________ of functional structures ________________________________ Ex: leg & hip bones in pythons & whales 5. Molecular Evidence Also called ______________________ evidence Compares biomolecules such as _______________________________between organisms Related organisms have ________________________________________________________________ How do new species form? 1. Geographic Isolation When members of a population are separated Ex: ___________________________________________________ 2. Reproductive Isolation When members of a population can’t breed even though they live nearby Ex: ____________________________________________________ 4 Different Types of Evolution 1. Divergent Evolution-_______________________ populations evolve ______________________________ Ex: __________________________________ changed independently _________________________________ 2. Convergent Evolution -Unrelated species become _____________ because they live in _______________________ Ex: shark & dolphin 3. Coevolution - Species that _____________________ adapt to _________________________ Ex: flowers & hummingbirds 4. Adaptive Radiation - Evolution of ___________________________ from _________________common ancestor Ex: ________________________________________ Population Genetics: Evolution at the Gene Level Population: _________________________________________________________________________________ _______________________________________________________________________________ Natural Selection: _______________________________________________________________________________ ________________________________________________________________________________ 4 Steps of Natural Selection: 1. 2. 3. 4. What is Gene Pool-----The gene pool is the set of all _________________________________, in any _____________________, usually of a particular species. This also proves to be the basic level at which _______________________________. Allele Frequency • The relative amount of an _________________________________ within a population (%) Example: If 100 individuals make up a population and 50 are homozygous dominant, 25 are heterozygous, and 25 are recessive, what is the allele frequency of the dominant allele? Your Turn: If 300 individuals make up a population and 150 are homozygous dominant, 100 are heterozygous, and the rest are recessive, what is the allele frequency of the recessive trait allele? How does natural Selection affect allele frequency? Mutations provide the raw material on which ________________________________ can act. Evolution depends on _____________ because this is the only way that _____________ among organisms are created Evolution acts on populations not ________________. Survival of the Fittest: If individuals having certain _________________ are better able to produce _____________________________ than those without them, the _____________________________________________ of those genes will increase. Survival: : ________________________________________________________________________ Fitness: : ________________________________________________________________________ Different Lines of Evidence: Wolf Lineage:_____________________________________________________________________________ Darwin’s Finches: ________________________________________________________________________ Genetic Drift: Mutations that take place because of random fluctuations of gene frequencies ___________________________________________________________________________ Example: __________________________________ -- a rare form of dwarfism that includes extra digits -- in the ______________________ population of eastern Pennsylvania, which has ___________________________ over many generations. Gene Flow: Changes in the _______________________ of a population because of the introduction of genes from _______________________________________________________. Example: ____________________________________was changed when U.S. Soldiers had children with Vietnamese women during the Vietnam war. Founder Effect: Changes in a population when a small population _________________________________________ bringing only a ___________________________________________________ and variation of the parent population. (The population then will contain only those genes the initial individuals brought with them) Example: __________________________________________ Bottleneck Effect: _________________________________________________________ leads to a population that no longer represents the original. Organisms are recessive at almost all alleles Mutations____________________________________________________________ Example: Cheetahs (2 bottleneck events) 1ST:10,000 years ago (_____________________) 2ND 1980’s ( _____________________________) (Population has rebounded from this population bottleneck to some 100,000 animals today. However, these animals are homozygous at every one of the gene loci that have been examined.) TYPES OF NATURAL SELECTION: • • • Stabilizing Selection: Extremes from ___________________ of the frequency distribution are _______________________. The most common form of natural selection. Example:___________________________________ Directional Selection: • Individuals at either end of the distribution are _____________________. The distribution shifts towards the ___________________________, depending on which one is favored. • What we usually think of as natural selection • Example: : __________________________________________________ ____________________________________________________________ Diversifying (Disruptive Selection) • Both _________________________________ at the expense of middle phenotype • A mechanism for ______________________________ without _________________________________ Cladistics • A Cladogram is a _______________________ that depicts species divergence from _______________________ • They show the ________________distribution and origins of ___________________________. • Cladograms are ______________________________________________________of phylogenetic relationships.