Akademik Veri Yönetim Sistemi
2nd FEMS Conference on Microbiology , Belgrade, Sırbistan, 30 Ağustos - 02 Eylül 2022, ss.538, (Özet Bildiri)
Background: S2 subunit mediating fusion between viral and host membranes. The high amino acid sequence identity of more than 90% in the S2 subunit suggests that the fusion mechanism during virus infection is well-conserved. With these properties, S2 is a better target for the development of a comprehensive therapeutic agent.
Objectives: The spike protein of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a major determinant of viral pathogenesis. Many researchers started to develop detection kits and anti-COVID-19 vaccines. It is aimed that production of target S2 subunit is required to facilitate the development of these tools.
Methods and Results: To enhance expression of the S2 subunit of SARS-CoV-2, S2 gene fragment (1788 base pair) was cloned in pET30a expression vector added a His tag at the C terminus. S2 subunit was expressed in Escherichia coli BL21 and Rosetta 2 pLysS cells induced by IPTG. The expression of S2 was optimized under different conditions including time, media, temperature etc. to obtain the proteins in soluble form. The bacterial cell pellet containing the recombinant proteins was lysed. IMAC purification was carried out over the S2 containing bacterial supernatant using the Akta Avant 25 chromatography system. The purified S2 protein (65.5 kDa) was confirmed by Western Blot using anti-polyhis antibodies. The resultant purified S2 subunit would be useful tools for the development of immunodetection kits, vaccines, and immunoglobulin production.
Key words: Covid-19 Antigen, Purification, Recombinant S2 Subunit, SARS-CoV-2.
*The present study was supported by The Scientific and Technological Research Council of TURKEY (TUBITAK) (Project number: 18AG020). We thank TUBITAK for financial support.