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Published byPosy Reynolds Modified over 9 years ago
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Liquid cultures to Lysis: Movie Clip from Teachers domain Student Guide Lab 6
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1.What is made of DNA? 2.What is made of protein? 3.How is this related to rfp?
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1.What is made of DNA? 2.What is made of protein?
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AraC dimer arabinose rfp Rfp protein rfp
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RFP Bruce Wallace
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RFP Bruce Wallace
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Growth of transformed bacteria on various plates P+ plates P- plates LBLB/ampLB/amp/ara LBLB/amp No growth Bruce Wallace
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The AraC Regulatory system
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PreLab 6 Questions Lab 5a: Questions -Pass cultures in the room -Expectations -Why they grew this way? - All about the AraC operon
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RFP expression araC generfp geneP BAD Transcription mRNA Translation araC protein Bruce Wallace
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RFP expression rfp geneP BAD araC protein araC gene araC protein prevents RFP transcription by causing a loop to form in the region of the fp gene r Bruce Wallace NO ARABINOSE:
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RFP expression araC protein arabinose araC generfp geneP BAD arabinose – araC protein complex RNA polymerase Arabinose – araC protein complex prevents DNA looping and helps to align RNA polymerase on the promoter site (P BAD ). mRNA Transcription Translation RFP (red fluorescent protein) Bruce Wallace With ARABINOSE:
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Colony isolation and culture Preparing an overnight culture of E. Coli for RFP expression LB/amp/ara broth Bruce Wallace
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Growth of transformed bacteria on various plates P+ plates P- plates LBLB/ampLB/amp/ara LBLB/amp No growth Bruce Wallace
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Purification of RFP from an overnight culture Overnight culture Cell pellet with RFP Lysed cells Pellet cell debris RFP with binding buffer Bruce Wallace
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