Incubation time optimization for the expression of recombinant CHIKV envelope protein E2 in the yeast Saccharomyces cerevisiae using SDS-PAGE

Abstract

Chikungunya virus (CHIKV) is the causative agent of Aedes-borne tropical disease Chikungunya. Clinical manifestations of Chikungunya are highly similar with other Aedes-borne tropical diseases, often making diagnosis by physical examination alone inaccurate. Differential diagnosis of Chikungunya is done through serological or molecular testing, both of which are costly and tedious. Effort has been made by Agency for the Assessment and Application of Technology (BPPT) to construct a CHIKV diagnostic kit based on the interaction of IgM and IgG patient against the envelope proteins of the virus. Hence production of CHIKV envelope protein is essentially needed to develop the kit. This process could be done by expressing the CHIKV envelope protein E2 in the yeast Saccharomyces cerevisiae INVSc1. The aim of this research is to optimize the expression of recombinant CHIKV envelope protein E2 in the yeast. Incubation time is the subject to change in the yeast expression system, in which protein will be detected and compared using SDS-PAGE protocol. Recombinant clones in expression medium are incubated each for 0 h, 24 h, 48 h and 72 h. Proteins are most expressed after 72 hours incubation time. Further analysis with western blotting is needed to detect the expressed protein E2 CHIKV.