question: The SARS-CoV-2 virus that caused pandemic COVID-19 has spread rapidly to humans and causes a very serious health and social problem around the world. In order to develop vaccine and therapeutic approaches, a comprehensive study of viral structure and viral biological infection needs to be understood. In addition to four structural proteins;spike glycoprotein (S), envelope (E), membrane protein (M), and nucleocapsid protein (N), SARS-CoV-2 viral genome, consist of two open reading frame (ORF) which is located in N-terminus, ORF1a, and ORF1b. The ORF1a consist of Main protease (3CLpro) and together with papain-like protease (PLpro) play a role in the cleavage of polyproteins and translated from viral RNA to mature protein. Here we describe a method of production of 6xHis-tagged Papain-like protease fragment in E. coli RIPL, including gene expression, solubilization of inclusion bodies by different methods, and detection of gene product target by Western Blot Analyses. © Published under licence by IOP Publishing Ltd.