Liquid cultures to Lysis:Movie Clip from Teachers domain

Student Guide Lab 6

1.What is made of DNA?2.What is made of protein?3.How is this related to rfp?

1.What is made of DNA?

2.What is made of protein?

AraC dimer

arabinose

rfp

Rfpprotein

rfp

RFP

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Growth of transformed bacteria on various plates

P+ plates

P- plates

LB LB/amp LB/amp/ara

LB LB/amp

No growth

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The AraC Regulatory system

PreLab 6 Questions

Lab 5a: Questions

-Pass cultures in the room-Expectations-Why they grew this way?

- All about the AraC operon

RFP expression

araC gene rfp genePBAD

Transcription

mRNA

Translation

araC protein

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RFP expression

rfp genePBAD

araC protein

araC gene

araC protein prevents RFP transcription by causinga loop to form in the region of the fp gener

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NO ARABINOSE:

RFP expression

araC protein

arabinose

araC gene rfp genePBAD

arabinose – araC proteincomplex

RNA polymerase

Arabinose – araC protein complex prevents DNA loopingand helps to align RNA polymerase

on the promoter site (PBAD).

mRNA

Transcription

TranslationRFP

(red fluorescentprotein)

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With ARABINOSE:

Colony isolation and culture

Preparing an overnight culture of E. Coli for RFP expression

LB/amp/arabroth

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Growth of transformed bacteria on various plates

P+ plates

P- plates

LB LB/amp LB/amp/ara

LB LB/amp

No growth

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Purification of RFP from an overnight culture

Overnightculture

Cell pelletwith RFP

Lysedcells

Pelletcell debris

RFP withbinding buffer

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