Successful expression of recombinant RNA dependent RNA polymerase (RdRp) of DENV-2 in E. coli cells.
Two step purification of enzymatically active RdRp protein using Ni-NTA and size exclusion columns.
High level production of RdRp with final yield of 6.5 mg protein from 1 L of culture.
Demonstration of in vitro polymerase activity of RdRp using homopolymeric RNA template.
Optimization of RdRp assay conditions such as protein concentration, GTP concentration and incubation time of the reaction.