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

PlasmidsPlasmids

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)

Polymerase Chain Polymerase Chain ReactionReaction

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

DNA FingerprintingDNA Fingerprinting

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

Selective BreedingSelective Breeding

Hybridization Ex: Burbank Potato

Inbreeding Ex: dogs

Transgenic OrganismsTransgenic Organisms

Bacteria --> human proteins

Mice --> immune genes

Livestock --> growth hormones

Plants --> genetically modified (GM)

Clone --> Dolly

Pre-Implantation Genetic Diagnosis

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