Plasmids  Bacterial DNA  Circular shape  Can be used to make proteins needed by humans

Steps of Plasmid Use 1.Plasmid is isolated and used as a vector 2.DNA from human obtained 3.Gene from DNA --> plasmid 1.Recombinant DNA - DNA that contains genetic information from more than 1 species 4.Transformation - process of inserting recombinant DNA back into bacteria 5.Cloning - making many copies of cell with recombinant DNA

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

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

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

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

PCR Steps 1.Denaturation 1.Heat DNA to separate 2.Annealing 1.Cool DNA so primers can attach 1.Primers are needed so polymerase can attach 3.Elongation 1.Polymerase adds on DNA nucleotides to make a copy of DNA (like replication) 1.taq polymerase 1.Special polymerase from bacteria in hot springs is used 2.Won’t denature during heating and cooling (steps 1-2)

Polymerase Chain Reaction

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

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

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

DNA Fingerprinting

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

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

Selective Breeding  Hybridization  Ex: Burbank Potato  Inbreeding  Ex: dogs

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

Pre-Implantation Genetic Diagnosis Pre-Implantation Genetic Diagnosis

References  Plasmid. (n.d.). Retrieved October 8, 2006, from Arizona State University Web site:  Simmer, M., & Secko, D. (n.d.). Restriction endonucleases. Retrieved October 8, 2007, from The Science Creative Quarterly Web site:  Gel. (n.d.). Retrieved October 8, 2006, from University of Miami Web site:  Southern blot. (n.d.). Retrieved October 8, 2006, from Towson University Web site:  Thermal cycler. (n.d.). Retrieved October 8, 2006, from images/Applied_Biosystems_GeneAmp_9700_96_Well_Thermal_Cycler.jpg  Polymerase chain reaction. (n.d.). Retrieved October 8, 2007, from  Russet Burbank potato. (n.d.). Retrieved October 8, 2007, from 8e.jpg