CHED 897 |
| West Nile virus (WNV) is a member of the flavivirus family that causes infection via viral transfer, typically from mosquitoes. The WNV genome encodes ten viral proteins, one of which is the NS5 protein, an RNA dependent RNA polymerase (RdRp). This RdRp is hypothesized to uniquely replicate the RNA genome of WNV during cell replication within a host cell. To investigate the kinetic properties of the RdRp, the NS5 gene was first cloned and then expressed with a C-terminal polyhistidine tail in E. coli and partially purified as a 100kDa protein using a cobalt affinity column. Enzymatic activities of the protein have been analyzed using a five part assay that incorporates radioactive ribonucleotides. The results presented here suggest that the WNV RdRp is time dependent, non-primer directed and prefers polyribonucleotide templates to oligoribonucleotide templates. Furthermore, initiation is greatly enhanced by the incorporation of an initial purine ribonucleotide. |
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Undergraduate Research Poster Session: Biochemistry
2:00 PM-4:00 PM, Monday, March 26, 2007 Hyatt Regency Chicago -- Riverside Center, Poster
Division of Chemical Education |