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Lecture 21

Biology 1002B Lecture 21: March 27

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Biology 1002B
Tom Haffie

March 27 How do you know youve been successful in getting your gene into another organism? Use reporter genes to confirm uptake GFP green fluorescent protein fluoresces green under UV light o Animals glow green because theyre expressing the reporter gene lacZ turns Xgal blue luciferase enzymes the fireflies use to make light Optogenetic construct the purple sequences (ITR) just terminal repeats: where the DNA gets integrated into the host chromosome EF1alphaiselongationfactortranslationfactorproducesverystronglyineverycell verystrong promoter o Grab the promoter from EF1 alpha ChR2 channelrhodopsin fused that with eYFP (yellow fluorescent protein) o There is an entire rainbow of fluorescent proteins o Both still work even though theyre fused together WPRE from a woodchuck virus gene 3UTR element that makes the message resistant to degradation so the message lasts longer hGH human growth hormone poly A signal Exome all of your sequences that are expressed Q: Which if the following characteristics of Taq polymerase makes it especially wellsuited for use in PCR reactions? Taq because its from thermos aquaticus (extremophile that functions in high temperatures) It is especially wellsuited because it is resistant to high temperatures PCR amplifies copy number of a specific sequence Denature the DNA add some specific primers these primers are DNA Prime DNA synthesis by the time at cycle 3, there are copies of the original product Copies increase exponentially to get dramatic amplification of specific sequences
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