Genetics: From Genes to Genomes

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Experiments generate model for regulation ofvirulence genes in V. cholerae.• Cloned two genes encoding subunits ofcholera toxin: ctxA and ctxB• Made ctxA-lacZreporter gene fusion• Created vector library of V. choleraegenomic DNA• Used E. coli to perform geneticmanipulationsCopyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display17-46
• Isolated a gene that regulates expression of ctx operon• Transformed E. coli containing ctx-lacZconstruct with clonescontaining V. cholerae DNA• Clones that contain a positive regulator should turn on ctx-lacZconstruct.• Identified ToxR, , a transmembrane protein• Identified ToxS, , helps ToxR form dimers which helps it bind to DNA.• What genes does ToxR regulate?• Gene fusions created to constitutive promoter• Fusion introduced into strains of V. cholerae with lacZ randomlyinserted around the genome• Identified intermediate regulator gene ToxT, , a transcriptional activatorthat binds to promoters of many genes, including ctx• ToxR/S or ToxT can activate the ctx genes that produce toxin• ToxT alone activates additional virulence genes which encode piliand other proteins.• Transcription of ToxT is regulated by ToxR/S.Copyright © The McGraw-Hill Companies, Inc. Permission required to reproduce or display17-47
Page 1 and 2: Powerpoint to accompanyGenetics: Fr
Page 3: Outline of Chapter 17• There are
Page 6 and 7: Fig. 17.2Copyright © The McGraw-Hi
Page 8 and 9: The utilization of lactose by E. co
Page 10 and 11: The Operon Theory• The players•
Page 12 and 13: The Operon Theory• Induction• L
Page 14 and 15: Experimental evidence for repressor
Page 16 and 17: Inducer releases repressor to trigg
Page 18 and 19: Changes in the operator can alsoaff
Page 20 and 21: Three experiments elucidate cis and
Page 22 and 23: ConstitutiveInsert Figure 17.9chere
Page 24 and 25: Some positive regulators increasetr
Page 26 and 27: Many DNA-Binding proteins containa
Page 28: Most regulatory proteins are oligom
Page 31 and 32: Using the lacZ gene as a reporter o
Page 33 and 34: Creating acollection oflacZinsertio
Page 35 and 36: The attenuation of gene expression:
Page 37 and 38: Termination of transcription finetu
Page 39 and 40: Global regulatory mechanisms coordi
Page 41 and 42: What enables transcription of σ 32
Page 43 and 44: Microarrays - a tool for uncovering
Page 45: Regulation of Virulence Genes inV.
Page 49: Unanswered Questions Remain• Why

Genetics: From Genes to Genomes

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