Plasmids Bacterial DNA Circular shape Can be used to make proteins needed by humans.
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Transcript of Plasmids Bacterial DNA Circular shape Can be used to make proteins needed by humans.
Steps of Plasmid UseSteps of Plasmid Use
1. Plasmid is isolated and used as a vector
2. DNA from human obtained
3. Gene from DNA --> plasmid1. 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 Inserting Gene into PlasmidPlasmid
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 - Testing for Gene - ElectrophoresisElectrophoresis
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 Testing for a Gene - Southern Blot- 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 Polymerase Chain Reaction (PCR)Reaction (PCR)
Makes copies of a gene in vitro (not in a living organism)
Done in thermal cycler
PCR StepsPCR 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)
Human uses of Genetic Human uses of Genetic Engineering - MedicalEngineering - 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 Human uses of Genetic Engineering - MedicalEngineering - 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 Human uses of Genetic Engineering - ForensicsEngineering - 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
Human Uses of Genetic Human Uses of Genetic Engineering - Engineering - AgricultureAgriculture
Genetically modified (GM) plants Can change:
Tolerance to drought Ripening speed Flavor Insect repellence Vitamin levels
Human Genome ProjectHuman 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
Transgenic OrganismsTransgenic Organisms
Bacteria --> human proteins
Mice --> immune genes
Livestock --> growth hormones
Plants --> genetically modified (GM)
Clone --> Dolly
ReferencesReferences 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