Expression of spotted duck interleukin-17 gene in E. coli

The duck interleukin-17 (duIL-17) gene was amplified by RT-PCR from the total RNA of conconavalin A (ConA)-stimulated spotted duck splenic lymphocytes. The gene was cloned into vector pCR2.1 to construct recombinant plasmid pCR2.1-duIL17. Sequencing result showed that duIL-17 had a silent mutation at 174 set from C to T, compared with the reference gene (EU366165.1). Then duIL-17 gene fragment was subcloned into the expression vector pGEX-6P-1, the resultant recombinant plasmid was transformed into E. coli BL21 and induced by IPTG, SDS-PAGE showed that the expressed fusion protein GST-duIL17 was about 45 ku. Western blot analysis showed that duIL-17 was recognized by chicken IL-17 antibody.