bacterial transformation by lexi chase colleen magowan
DESCRIPTION
Restriction Enzymes ● Cuts plasmid at one specific sequence ● Creates sticky endsTRANSCRIPT
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Bacterial Transformation
By Lexi Chase & Colleen Magowan
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Stanley Cohen and Herbert Boyer ● Studied E. Coli PSC101 gene
● Transfers antibiotic resistance● Also transferred frog genes to
bacteria
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Restriction Enzymes
● Cuts plasmid at one specific sequence
● Creates sticky ends
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LigaseGlue plasmid & old DNA
together: creates recombinant DNA
Antibiotic resistant plasmid was being created
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Heat Shocking the E. Coli…● Creates a pressure
difference between inside and outside of the cell
● Forms pores--->new genes (from plasmid) get in
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How did they find it?● DNA sequencing WITHOUT COMPUTERS● Lots of guess and check● By hand, they searched for repetition and
patterns within the human genome (1,000’s of genes)
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Results
● Bacteria that grew in the presence of both antibiotics contained the recombinant DNA
● Bacteria that only grew on one contained the old plasmids
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Yes!Cohen & Boyer went on to add toad genes
to bacteriaLater generations of bacteria kept making
the toad proteins from the genesInterspecies gene swaps are possible
Can we do this with human genes?
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Heat shocking: is it worth it?
Yes!
● Less efficient without heat shocking
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Sourceshttp://www.genomenewsnetwork.org/resources/timeline/1973_Boyer.php
http://www.chemheritage.org/discover/online-resources/chemistry-in-history/themes/pharmaceuticals/preserving-health-with-biotechnology/berg-boyer-cohen.aspx
https://www.google.com/search?q=fire&rlz=1C1CHWA_enUS626US626&source=lnms&tbm=isch&sa=X&ved=0ahUKEwisnq3x59XKAhXGph4KHfZQC2AQ_AUIBygB&biw=1366&bih=599#tbm=isch&q=fire+clipart&imgrc=I74Li8iHvTBcgM%3A
https://en.wikipedia.org/wiki/African_clawed_frog
http://parts.igem.org/Part:BBa_K338001
http://www.bville.org/teacherpage.cfm?teacher=427