Evaluation I.Key

The document discusses cloning, expressing, purifying, and characterizing the NS2B/NS3 protease/RNA helicase protein of Japanese encephalitis virus. Specifically, it aims to clone the NS2B-NS3 portion of the JEV polyprotein and express it in E. coli to study its cleavage activity and cofactor requirements compared to dengue virus. It also aims to analyze differences in substrate specificity and inhibition using fluorogenic or chromogenic peptide substrates. The results show successful amplification and cloning of the NS2B-NS3 region from JEV as well as expression and purification of the NS2B-NS3 protease for further enzymatic characterization.