RBD purification of  spike 2 SARS-COV expressed gene  in prokaryotic cell and investigation of antiviral peptides binding with it using bioinformatics studies

Sadeghizadeh, Majid; Moazzezi Tehrankhah, Narges

RBD purification of spike 2 SARS-COV expressed gene in prokaryotic cell and investigation of antiviral peptides binding with it using bioinformatics studies

Document Type : Original Research

Authors

Majid Sadeghizadeh ¹

Narges Moazzezi Tehrankhah ²

¹ Professor of Genetics, School of Biological Sciences, Tarbiat Modares University.Tehran/Iran

² Department of Genetics, Faculty of Biological Sciences, Tarbiat Modares University

Abstract

The countries’ social and economic conditions have been threatened by corona epidemic and a large number of people’s death in the world. SARS-CoV-2 virus, a form of corona virus family, is responsible for corona disease and its spread in the present century. The study of the receptor binding region (RBD) in the spike protein is very important for scientists because the new corona virus uses its surface spike protein for binding to the ACE2 surface protein and entering its genetic material to the host cells. By this protein and its receptor binding region inhibition, the prevention of virus entrance to the cell is possible. The virus’s genes can be multiplied by cloning and its protein can be purified. The usage of antiviral peptides as the most practical methods and binding inhibitory peptides of RBD to the ACE2 receptor for SARS-CoV-2 treatment, are of great interest to scientists. In the present research, RBD cloning in PET28a expression vector, RBD protein expression and GFP/RBD fusion protein were performed in prokaryotic host. Due to this protein’s insolubility in the prokaryotic host, column refolding was performed with urea gradient with a nickel-agarose column and the synthesized protein was confirmed through western blot technique. Three nominated peptides from articles used to compare their binding to RBD using bioinformatics and their tendency to bind to each other was investigated by molecular docking. The mentioned peptides can be used in this virus infection treatment due to their binding potential to RBD, if their interaction is proven.

Keywords

SARS-CoV-2

RBD

Refolding

Antiviral peptides

Subjects

Bioinformatics

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