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TRYP and LAC OPERON SIMULATION Name: ___________________________________________ Period: ____ Scoring: Part 1: ____ / 2 drawings + ____/ 3 Questions = ______/ 5 Part 2: _____ / 3 drawings + ____/ 6 Questions = ______/ 9 Quiz: _____/ 14 Total: _____/ 28 Part 1: TRYP OPERON SIMULATION LAB 1)[ tryp] = high => gene activity (low or high) Promoter Operator Repressor Binding Site Tryp E gene Tryp D gene 2)[tryp] = low => gene activity (low or high) Tryp C gene Part 1 QUESTIONS: 1) Why is the tryp operator described as repressible? 2) For repressible operons, is the gene activity changed in response to the presence of the precursor in the pathway or the final product metabolite in the pathway? 3) How does a repressor prevent transcription? Part 2: LAC OPERON SIMULATION DRAWINGS QUESTION # 1: Circle the correct choice: If [Glucose] = high, then [cAMP] = high or low Conditions to simulate: 1) [Glucose] = high, [lactose] = low PROMOTER Operator CRP or CAP Site lacZ gene lacY gene lacA gene 2) [Glucose] = high, [lactose] = high PROMOTER CRP or CAP Site Operator lacZ gene lacY gene lacA gene 3) [Glucose] = low, [lactose] = high PROMOTER Operator CRP or CAP Site lacZ gene lacY gene lacA gene Part 2 Questions to answer: 2) Is the lactose the precursor or the final product of the metabolic pathway for the lac genes? 3) Why is the lac operon described as inducible? 4) Describe how a high [cAMP] activates transcription. 5) Given the following pathway X -> Y -> Z, A) If [X] primarily turns the gene on and off, the pathway is most likely to be (inducible or repressible) B) If [Z] primarily turns the gene on and off, the pathway is most likely to be (inducible or repressible) Extension: Genetic Studies of the Lac Operon – BE SURE TO READ BACKGROUND INFORMATION IN LAB MANUAL FIRST! Discuss the data with your lab team but make sure your final answers are your own individual. This question is not a part of operon lab quiz) EXPLANATION OF CLASS I MUTANTS: 1) Below is a diagram of the lac operon model proposed by Jacob and Monot. Note: I gene codes for repressor protein. Genes Z, Y and A code for proteins directly involved in bringing lactose into the cell and breaking it down. (Note: Z is the gene for beta-galactosidase; recall that the activity of this enzyme is used to monitor operon expression.) A) There are three possible regions of DNA mutation: 1) The I gene that codes for repressor protein. 2) The operator region 3) The lactose metabolism genes (Z,Y,A) Based on the data from Table 2 which one of these regions can be immediately eliminated and why? B) On the diagram below, CIRCLE the location of the mutation. Mutated Lac Operon on Chromosome I gene Operator Z gene Y gene A gene Proteins for Lac Digestion, including beta-galactosidase Repressor Protein Wild Type copy Lac Operon on Plasmid I gene Operator Z gene Y gene A gene Repressor Protein C) Using symbolic pictures for repressor protein and/or notations on the diagram as well as words explain how the mutation you propose is consistent with the experimental data. EXPLANATION OF CLASS II MUTANTS: 2) Below is a diagram of the lac operon model proposed by Jacob and Monot. Note: I gene codes for repressor protein. Genes Z, Y and A code for proteins directly involved in bringing lactose into the cell and breaking it down. (Note: Z is the gene for beta-galactosidase; recall that the activity of this enzyme is used to monitor operon expression.) A) There are three possible regions of DNA mutation: 1) The I gene that codes for repressor protein. 2) The operator region 3) The lactose metabolism genes (Z,Y,A) Based on the data from Table 2 which one of these regions can be immediately eliminated and why? B) On the diagram below, CIRCLE the location of the mutation. Mutated Lac Operon on Chromosome I gene Operator Z gene Y gene A gene Proteins for Lac Digestion, including beta-galactosidase Repressor Protein Wild Type copy Lac Operon on Plasmid I gene Operator Z gene Y gene A gene Repressor Protein C) Using symbolic pictures for repressor protein and/or notations on the diagram as well as words explain how the mutation you propose is consistent with the experimental data.