Expression and purification of MERS-CoV envelope protein, an essential viroporin, using the baculovirus expression system

Alsaadi, E., Alghezi, D. and Jones, I. ORCID: https://orcid.org/0000-0002-7738-2516 (2023) Expression and purification of MERS-CoV envelope protein, an essential viroporin, using the baculovirus expression system. Iranian Journal of Microbiology, 15 (1). pp. 121-127. ISSN 2008-4447 doi: 10.18502/ijm.v15i1.11926

Abstract/Summary

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, recombinant 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.

Item Type	Article
URI	https://reading-clone.eprints-hosting.org/id/eprint/111823
Identification Number/DOI	10.18502/ijm.v15i1.11926
Refereed	Yes
Divisions	Life Sciences > School of Biological Sciences > Biomedical Sciences
Uncontrolled Keywords	MERS-CoV, Insect cells, Coronaviruses, Middle east respiratory syndrome, Baculovirus, Envelope protein, Immobilised metal-affinity chromatography
Publisher	Teheran University of Medical Sciences
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