expression and purification of mers-cov envelope protein, an essential viroporin, using the baculovirus expression system

Background and objectives: the causative agent of middle east respiratory syndrome (mers) is a zoonotic coronavirus (mers-cov) identified in saudi arabia in 2012. the envelope (e) protein of mers-cov is a small viral protein which plays several essential roles during virus replication. to facilitate study of the structure and function of the e protein, recom- binant mers-cov e protein was expressed using the baculovirus expression system. materials and methods: a recombinant e open reading frame including an 8-histidine tag at the amino terminus was designed and cloned into a baculovirus transfer vector. following construction of a recombinant virus insect cells were infected and the expression of the e protein assessed by sds-page and western blotting.results: recombinant e protein, tagged at the n-terminus with a polyhistidine sequence, with a molecular mass of 10.18 kd was identified by western blotting with an anti-his antibody. following large scale infection e protein was released by detergent mediated lysis of infected cells and purified by immobilized metal ion affinity chromatography (imac). conclusion: purified full length recombinant mers-cov e protein can be isolated by imac and is suitable for further functional, biophysical or immunological studies.