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- Describe the p-nitrophenol assay and how a standard curve can be used to calculate the concentration of an unknown .As shown , several medical agents are now commercially produced by genetically engineered microorganisms. Discuss the advantages and disadvantages of making these agents this way.What reaction would you expect when performing a positive control in the catalase assay? What would it mean if a known catalase-positive bacteria did not produce the expected reaction?
- Give an example for any biopharmaceutical produced by Recombinant DNA Technology (except insulin). What is the name of the drug/trademark? Write its mode of action. List the experimental steps under the laboratory conditions List the production steps in the factoryWhen Avery and his colleagues had obtained what was concluded to be the transforming factor from the IIIS virulent cells, they treated the fraction with proteases, RNase, and DNase, followed in each case by the assay for retention or loss of transforming ability. What were the purpose and results of these experiments? What conclusions were drawn?Why is the DNS method used in glucose assay?
- If a concentrated sample is out of the detection range of a chosen dye-based assay, what can be done?How would you determine the concentration of a protein that falls outside (above or below) the linear range of this assay? asapYou have isolated a new endoglucanase enzyme. Through various enzymes assays, you are trying to determine the optimal conditions for this enzyme's activity. You make the following observations: i. when performing the assay at 40°C using buffers with different possibilities, you observe that the enzyme has no activity below pH 3 or above pH 11. You observe high activity at pH 6 and about 50% of this activity level at pH 5 and pH 7. ii. When performing the assay using a buffer at pH 6, you measure almost no activity at a temperature of 10°C. However, the activity steadily rises when tested at 20°C, 30°C, 40°C and 50°C, where the highest activity was measured, before declining at 60°C. At 70°C there is no measurable activity. Explain the meaning of these observations and identify the optimal pH and temperature conditions for the endoglucanase enzyme. Further more, explain why these observations occur based on your knowledge of the effects of pH and temperature on enzymes.