| One of the main goals of the research project was to purify and characterize the SARS coronavirus RNA-dependent RNA polymerase. We recently achieved a major breakthrough with the purification and initial characterization of this enzyme. Our main focus is now to further characterize this enzyme biochemically and its molecular collaboration with other crucial viral replication enzymes. Most of these enzymes have already been purified in our lab and we foresee significant progress in this direction in the next two years. Additionally, in our present research goals we include now single molecule studies and the characterization of the recently purified SARS coronavirus RNA-dependent RNA polymerase (RdRp) and accessory enzymes (e.g. the primase and helicase) at this level using a magnetic tweezers setup. This approach should enable us to characterize the enzyme kinetics under a large range of parameters at the single molecule level and it should thus help us to create a more detailed description of the enzyme?s function. Initial experiments have already been performed in collaboration with Prof Dr Nynke H. Dekker at the TU Delft and are currently being optimized. In conclusion, our initial research plans are still viable and we have already made significant progress towards reaching our main research goal. We have a good experimental foundation for the remainder of the grant and have even extended our initial research goals with in dept single molecule studies. |
