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TAMU BIOL 111 - Gene Expression Lab

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Name: Chloe EzellGroup: 2Gene Expression LabLab Activity 1: Table 1CultureConditionsRxn#Lysis ReagentE. coliCulture VolumeUnusedmedia fordifferencesin enzymeZ buffer to bring to2 mL totalVol. ONPG(4 mg/mL)for 0.8 mgLB only(plain media)1a 20 µL 20 µL 180 µL1380 uL400 uL1b 20 µL 40 µL 160 µL1380 uL 400 uL1c 20 µL 200 µL 01380 uL 400 uLLB+lactose(lactose culture)2a20 µL20 µL180 µL1380 uL400 uL2b 20 µL 40 µL 160 µL1380 uL 400 uL2c 20 µL 200 µL 01380 uL 400 uLLB+glucose(glucose culture)3a20 µL20 µL180 µL1380 uL400 uL3b20 µL40 µL 160 µL1380 uL 400 uL3c20 µL200 µL 01380 uL 400 uLLB+glucose+lactose (glucose+lactoseculture)4a20 µL20 µL180 µL1380 uL400 uL4b20 µL40 µL 160 µL1380 uL 400 uL4c20 µL200 µL 0 1380 uL 400 uLBLANKBlank20 µL0 µL 200 µL1380 uL 400 uLLab Activity 1: Table 2Rxn # Time started Time stopped Elapsed time (sec) A420nmActivity(A420nm/time)*10001a0:00 43:15 2595 0.02 0.00771b0:15 38:45 2315 0.06 0.02591c0:25 26:55 2590 0.21 0.13212a0:50 34:20 2010 0.009 0.00452b1:00 26:35 1535 0.17 0.11072c1:10 17:40 990 0.60 0.60613a1:40 43:30 2510 0.01 0.00403b1:50 39:10 2220 0.08 0.03573c2:00 7:15 1515 0.24 0.15844a2:30 43:45 2475 0.01 0.00404b2:40 39:30 2210 0.13 0.05884c2:50 27:35 1485 0.27 0.18181Name: Chloe EzellGroup: 2Lab Activity 2: Table 3.1Table 3.2Tube 1CultureConditionsRxn#LB broth fordifferences inenzymeLysis ReagentE. colivolumefrom Tube 2Z buffer tobring to 2 mLtotalVol. ONPG(4 mg/mL) for0.8 mgLB culture + Lactose1 80 µL 10 µL 20 µL1690200Glucose culture+Lactose2 80 µL 10 µL 20 µL1690200LB culture + Glucose380 µL10 µL20 µL1690200Lactose culture+ Glucose480 µL10 µL20 µL1690200BLANK2Tube 2CultureConditionsRxn#µL 20%LactoseµL20% GlucoseE. coli cultureLB culture+Lactose1 50 µL 0 µL 950 µLGlucose culture+Lactose2 50 µL 0 µL 950 µLLB culture+Glucose30µL 50 µL 950 µLLactose culture+ Glucose4 0 µL 50 µL 950 µLBLANKName: Chloe EzellGroup: 2Table 4Rxn # Time started Time stopped Elapsed time (sec)A420nmActivity(A420nm/time)*100010:10 1:44:59 94.059 0.03 0.318920:10 29:20:00 1750 0.00 0.000030:10 52:10:00 3120 0.01 0.003240:10 26:10:00 1560 0.09 0.0577Create a graph of your results for Lab Activity 1. Plot enzyme activity vs enzyme concentration (a, b, c) for each treatment. The graph should include a title, labeled axes, and labeled data lines.1. Under what conditions is the LacZ gene expressed? Is there any sign of activation or repression? Be sure to refer to your results when discussing your answer. The LacZ gene is expressed when lactose is present and lac operon is started. Repression occurs as the volume decreases, while activation happens when the volume increases. Allolactose binds to the protein, which activates it.2. Examine your data for Lab Activity 2. Describe the data and your interpretation of what you see.Based on the data from Lab Activity 2, sugar does have an interaction with the assay. The allolactose found in E.Coli binds to the protein instead of the ONPG, which decreases the activity rate. The lac operon deactivates in the presence of glucose, which causes the repression. This can be seen in table 4, 3Name: Chloe EzellGroup: 2which shows the initial activation, then repression due to the sugars, then slight increase in activity once the sugar has been


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