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All Journal HAYATI Journal of Biosciences
Yuliana
Primate Research Center, IPB University, Bogor 16151, Indonesia
Agriculture, Biological Sciences & Forestry Earth & Planetary Sciences

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Expression of SARS-CoV-2 Nucleocapsid (N) Recombinant Protein Using Escherichia coli System Rizki Aulia Ansari; Uus Saepuloh; Silmi Mariya; Yuliana; Rachmitasari Noviana; Irma Herawati Suparto; Huda Shalahudin Darusman
HAYATI Journal of Biosciences Vol. 30 No. 3 (2023): May 2023
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.30.3.445-450

Abstract

One of the main antigen that can be used for serological testing is the nucleocapsid (N) which is the most abundant viral-derived protein in SARS-CoV-2 where this virus can cause COVID19 disease. The aim of this study was to develop the SARS-CoV-2 N recombinant protein using Escherichia coli expression system. A total of 1,089 nucleotides encoding 362 amino acids of SARS-CoV-2 N was cloned to pET-14b vector. The plasmid then expressed in E. coli BL21 (DE3) and induced with 1.0 mM IPTG (Isopropyl-β-d-1-thiogalactopyranoside). The cell was harvested using denaturation lysis buffer due to inclusion body formation of SARS-CoV-2 N protein. Dialysis processed and concentrated using PEG-6000 resulted 0.992 mg/ml protein yield. Analysis of SARS-CoV-2 N recombinant protein using SDS-PAGE technique showed approximately 37.0 kDa specific band target protein. Application of this SARS-CoV-2 N recombinant protein to vaccinated and non-vaccinated antibody serum samples using ELISA technique indicated the significant result of optical density mean at 0.603 and 0.135, respectively. This study revealed that the production of SARS-COV-2 N recombinant protein could be carried out in E. coli expression system under denatured conditions, therefore the methods are more effective in producing the protein as a basic material in immuno-diagnostic assay.
Co-Authors Huda Shalahudin Darusman Irma Herawati Suparto Rachmitasari Noviana Rizki Aulia Ansari Silmi Mariya Uus Saepuloh