Bakteri ve mayalarda SARS-COV-2 spıke geninin klonlanması ve ifade edilmesi

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is an enveloped, positive-sense, single-stranded RNA virus that causes coronavirus disease (COVID-19) that emerged in 2019 in Wuhan city of China. Virus particles contain an RNA (mRNA), structural proteins, non-structural proteins (NSPs), and accessory proteins. After the virus enters the cell via receptors on the surface of host cells, 4 structural proteins; Spike, Membrane, Nucleocapsid, Envelope (S, M, N, E), about 20 non-structural proteins (NSPs), and a dozen accessory proteins (APs) are made. While structural proteins are required for virus entry, assembly, and immune evasion, NSPs and APs are important in transcription, replication, and host cell control. Since the Spike glycoprotein, the largest protein of the virus, mediates viral attachment and entry into the host, it has been an important target for vaccine development, monoclonal antibody therapies, and antigen-based or PCR diagnostic tests. In this study, the Spike gene was cloned into a shuttle vector that can replicate and express in both bacteria and yeast, and the expression profile of the protein was investigated. Our study showed that both bacteria and yeast expressed the Spike protein and the recombinant shuttle vector was stably maintained in both cell types. Keywords: SARS-CoV-2, COVID-19, Coronaviruses, Spike protein, Shuttle vectors, Escherichia coli, Saccharomyces cerevisiae.