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-dulL17 was about 45 ku. Western blot analysis showed that duIL-17 was recognized by chicken IL-17 antibody.%为表达麻鸭白介素17(duIL-17),本研究提取ConA刺激的麻鸭脾脏淋巴细胞总RNA,RT-PCR扩增duIL-17基因片段,将该片段插入克隆载体pCR2.1中构建重组质粒pCR2.1-duIL17.将duIL-17基因片段亚克隆到原核表达载体pGEX-6P-1中,构建重组质粒pGEX-duIL17,将其转化到大肠杆菌BL21中,IPTG诱导表达.测序结果显示,duIL-17与参考序列(EU366165.1)相比较第174位碱基由C突变为T,并且为沉默突变.SDS-PAGE和western blot分析显示,诱导表达的重组蛋白GST-duIL17主要以包涵体形式存在,分子量大小约为45 ku,duIL-17与抗鸡IL-17抗体具有良好反应性.本研究为制备duIL-17单克隆抗体提供了实验依据.
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