Wednesday, February 26, 2020
Molecular basis of RNA-dependent RNA polymerase II activity Coursework
Molecular basis of RNA-dependent RNA polymerase II activity - Coursework Example The RdRP activity has also been implicated in the replication of the RNA Genomes of HDV virus and plant viriods. 3) What is meant by HDV? Why would investigators want to study HDV? In your answer, tell me about HDV and why investigators would want to study HDV. What is the ANTIGENOME? Why is it important in the experimental design? HDV is Hepatitis Delta Virus. The investigators have taken HDV for the study as RdRP activity of Pol II has been established to be involved in the replication of the viral genome. Antigenome is the complementary positive RNA which serves as the template for the synthesis of the negative strand of the virus. To understand the physiological significance of the RdRP activity, the terminal segment of the HDV antigenome, which directs RNA synthesis and is sensitive to the Pol II inhibitor alpha- amanitin, was studied. 4) What is meant by Ã¢â¬Å"FAM-labeled RNA products were separated by denaturing gel electrophoresisÃ¢â¬ ? DO NOT give me cookbook recipes, i.e. do not provide a Ã¢â¬Ëstep-by-step how toÃ¢â¬â¢ that could be used to perform an experiment. Describe the method in general terms. FAM is 6 carboxy- flourescein label. Nucleotides labeled with such flourophores, have shown improved sensitivity, versatility and normalization when separated by denaturing gel electrophoresis. After purification of the nucleotides the fluorescein tag was attached by incubation with the 5Ã¢â¬â¢ terminal of the nucleotides. 5) DATA INTERPRETATION: Evaluate Figure 1. Break the analysis down. Strive to understand what each lane of the gel represents. Make note of the controls as well. Your focus for your response is the part of the figure labeled b, c, and d. Use part a in assisting you. Summarize what each part (b, c, and d) of the figure suggests. The RNA synthesized by the RdRP activity of Pol II is fluorescent labeled and separated by gel electrophoresis and viewed. Lane 1 in the gel shows only the reactant RNA.