Abstract: In this study, the DNA encoding sequences of Cpf1 nuclease fragment (N-terminal 166 amino acids) were subcloned into pET-28a (+) vector and expressed in E. coli BL21. The IPTG concentration and induction time were optimized, and the purified protein was used to immunize New Zealand white rabbits. The titer of antiserum reached 128 000 by indirect ELISA method. The antibody was purified by Protein A resin and identified by Western Blot. The results showed that Cpf1(1-166) and Cpf1 expressed in E. coli could be specifically identified. This antibody provids a tool for the detection of Cpf1 in organisms and lay a foundation for promoting the application of CRISPR/Cpf1 system.

Key words: CRISPR-Cpf1, Escherichia coli, prokaryotic expression, protein purification, polyclonal antibody