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Unformatted text preview: Sarah Song March 25, 2008 BIO 205.33 Lab 20 Bacterial Transformation Notes Lab I ntroduction • Gene cloning: technique for making copies of desirable genes o Isolate a gene from one organism and transfer to another Desirable genes inserted into plasmids • Transformed cell: modified DNA inserted into host cell • Antibiotic resistance separated transformed cells from normal cells • Plasmids are replicated rapidly o Many copies of inserted genes produced in transformed bacterial cell High level of expression, host bacteria produces large amounts of foreign protein I ntroduction to Genetic T ransformation • Genetic transformation: insertion of a gene into an organism in order to change the organism’s traits • Genetic transformation in biotechnology: o Agriculture: frost, pest, or spoilage resistance in plants o Bio-remediation: bacteria able to digest oil spills o Medicine: sick person’s cells transformed with healthy copies of the diseased gene • Plasmid: small, circular pieces of DNA that enter the bacterial cytoplasm o Usually contain genes for traits beneficial to bacterial survival o Bacteria can transfer plasmids back and forth to share beneficial genes Resistance to antibiotics • Genetic engineering: genes that code for new traits can be inserted into a plasmid • pGLO plasmid o Genetic features: GFP Green Flourescent Protein • Glow in the dark gene taken from the bioluminescent jellyfish Aequorea vitoria • Stimulated by pathways specific to jellyfish o E. coli will not be able to glow on their own o Shine UV light on GFP to see emission of green color Ability to resist the antibiotic ampicillin (antibiotic) “DNA switch” • Gene regulation system used to control the expression of GFP • pBAD and araC promoter Sarah Song March 25, 2008 BIO 205.33 Lab 20 Bacterial Transformation o Enables t ranscript ion of any gene it is attached to, if arabinose...
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This note was uploaded on 10/13/2008 for the course BIO 205 taught by Professor O'neal during the Spring '08 term at SUNY Stony Brook.
- Spring '08