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Genetic Engineering

Genetic Engineering. Biology CPE. Plasmids. Bacterial DNA Circular shape Can be used to make proteins needed by humans. Steps of Plasmid Use. Plasmid is isolated and used as a vector DNA from human obtained Gene from DNA --> plasmid

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Genetic Engineering

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  1. Genetic Engineering Biology CPE

  2. Plasmids • Bacterial DNA • Circular shape • Can be used to make proteins needed by humans

  3. Steps of Plasmid Use • Plasmid is isolated and used as a vector • DNA from human obtained • Gene from DNA --> plasmid • Recombinant DNA - DNA that contains genetic information from more than 1 species • Transformation - process of inserting recombinant DNA back into bacteria • Cloning - making many copies of cell with recombinant DNA

  4. Inserting Gene into Plasmid • Restriction enzymes cut DNA /plasmid • Makes sticky ends • Allows for overlap site for DNA and plasmid to attach to one another • Gene inserted into plasmid • Ligase = paste human DNA and bacterial plasmid together • Result = recombinant DNA

  5. Testing for Gene - Electrophoresis • Recombinant Plasmid removed and broken down • Plasmid pieces --> wells in agarose gel • Agarose has holes that DNA pieces travel through • Charged field pulls (-) charge DNA through gel • Small pieces move fast • Large pieces move slow • Results in banding

  6. Testing for a Gene - Southern Blot • DNA bands --> nitrocellulose filter paper • Radioactive probes are added • Attach to inserted gene • X-Ray Radiography - taking a picture to find radioactive probes attached to inserted gene

  7. Polymerase Chain Reaction (PCR) • Makes copies of a gene in vitro (not in a living organism) • Done in thermal cycler

  8. PCR Steps • Denaturation • Heat DNA to separate • Annealing • Cool DNA so primers can attach • Primers are needed so polymerase can attach • Elongation • Polymerase adds on DNA nucleotides to make a copy of DNA (like replication) • taq polymerase • Special polymerase from bacteria in hot springs is used • Won’t denature during heating and cooling (steps 1-2)

  9. Polymerase Chain Reaction

  10. Human uses of Genetic Engineering - Medical • Production of Medicine • Ex: insulin for diabetes • Vaccines • Solution containing all or part of a harmless version of pathogen • Immune system makes antibodies for pathogen --> next time body is exposed it is prepared to fight the pathogen

  11. Human uses of Genetic Engineering - Medical • Gene Therapy • Healthy copy of a gene is put into the cells for a person whose gene is defective • The new cells produce the missing protein

  12. Human uses of Genetic Engineering - Forensics • DNA fingerprinting • all people (minus identical twins) have different DNA sequences • When exposed to restriction enzymes different sequences get cut in different locations • Causes different bands when electrophoresis is done • Used to identify individuals in crimes, when biological material is left at the scene

  13. DNA Fingerprinting

  14. Human Uses of Genetic Engineering - Agriculture • Genetically modified (GM) plants • Can change: • Tolerance to drought • Ripening speed • Flavor • Insect repellence • Vitamin levels

  15. Human Genome Project • Used to determine the nucleotide sequence of the entire human genome and map the location of every gene on each chromosome • Started in 1990 and completed in 2003

  16. Selective Breeding • Hybridization • Ex: Burbank Potato • Inbreeding • Ex: dogs

  17. Transgenic Organisms • Bacteria --> human proteins • Mice --> immune genes • Livestock --> growth hormones • Plants --> genetically modified (GM) • Clone --> Dolly

  18. Pre-Implantation Genetic Diagnosis

  19. References • Plasmid. (n.d.). Retrieved October 8, 2006, from Arizona State University Web site: http://www.west.asu.edu/lsi/sum04/wk4/page3.htm • Simmer, M., & Secko, D. (n.d.). Restriction endonucleases. Retrieved October 8, 2007, from The Science Creative Quarterly Web site: http://www.scq.ubc.ca/?p=249 • Gel. (n.d.). Retrieved October 8, 2006, from University of Miami Web site: http://www.bio.miami.edu/dana/250/25002_10.html • Southern blot. (n.d.). Retrieved October 8, 2006, from Towson University Web site:http://pages.towson.edu/cberkowe/medmicro/images?D=A • Thermal cycler. (n.d.). Retrieved October 8, 2006, from http://www.labcentraal.com/webinventory/ images/Applied_Biosystems_GeneAmp_9700_96_Well_Thermal_Cycler.jpg • Polymerase chain reaction. (n.d.). Retrieved October 8, 2007, from http://athena.nhaminated.com/images/pcr2.jpg • Russet Burbank potato. (n.d.). Retrieved October 8, 2007, from http://www.inspection.gc.ca/english/plaveg/potpom/var/russetburbank/potato28e.jpg

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