Overexpression of recombinant domain III envelope protein of Zika virus

Aims: Zika virus (ZIKV) is a member of the Flaviviridae family and is transmitted to humans by mosquitoes. In humans, it causes disease known as Zika fever. The severity of the infection ranged from asymptomatic to mild disease and to infection associated with neurological disorders and congenital anomaly. The common symptoms are maculopapular rash, fever, arthralgia, myalgia, headache and conjunctivitis. The flavivirus genome consists of structural and nonstructural proteins. The envelope (E) glycoprotein is the major structural protein which is responsible for virus entry and represents a major target for neutralizing antibodies. The E protein consists of three distinct domains: domain I, domain II and domain III. The domain III (DIII) of the E protein has shown to be useful as antigen for flavivirus serologic diagnosis and immunization in animal model. Hence, the aim of this work is to express the DIII of E protein (EDIII) of ZIKV for immunoreactivity study. Methodology and results: The EDIII of ZIKV was cloned into pET SUMO cloning vector and transformed into Mach-T1 competent E. coli cells. Positive clone was selected, verified and transformed into BL21 (DE3) competent E. coli for protein expression. The expression of the recombinant protein was analysed on SDS-PAGE and western blot. The recombinant fusion protein of EDIII/SUMOHIS (rEDIII) was successfully expressed at a molecular weight of approximately 38.2 kDa. Conclusion, significance and impact of study: The expression of the protein was confirmed by detection with anti-histidine and a flavivirus antiserum, HPR.