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Session 6, Abstract 37

EPITOPE TAGGING AND FUNCTIONAL ANALYSIS OF THE TuMV REPLICATION PROTEIN

Ziomara Jurado* and Garcia-Ruiz (Jerald Bricker), Biology Department, Nebraska Wesleyan University, Lincoln, NE 68504; Department of Plant Pathology, University of Nebraska-Lincoln, Lincoln, NE, 65583

RNA-dependent RNA polymerase (RdRp) is used by RNA viruses to replicate a model plant virus. The RdRp is aided by other viral and host proteins. For the Turnip Mosaic Virus (TuMV), NIb is the RdRp. The viral and host proteins that assist NIb in replication are currently unknown. In order to perform immunoprecipitation to identify the unknown proteins, epitope tagging of NIb was done using two different tags, alone or in combination: 3xFLAG and 6xHIS-3xFLAG. Nicotiana benthamiana and Arabidopsis thaliana were inoculated with the WT and tagged TuMV to measure the viral accumulation. The NIb-tag was functional within the virus and it does replicate similar to the wild-type (WT), except it has a slower infectivity rate. Epitope-tagged NIb will be used to identify viral and host proteins that aid NIB with viral replication by immunoprecipitation and proteomic approaches.