目的克隆、表达十二指肠钩虫巨噬细胞迁移抑制因子(MIF)AduMIF-1基因。方法设计、合成特异引物,以十二肠钩虫成虫cDNA为模板,通过PCR扩增AduMIF-1基因。将获得的AduMIF-1编码序列克隆至原核表达载体pET32a,构建重组表达质粒pET32a/AduMIF-1。重组表达质粒转入大肠埃希菌BL21(DE3)中,IPTG诱导表达并分离纯化重组AduMIF-1。结果成功扩增到AduMIF-1全长编码序列,完整阅读框长度为360bp,编码119个氨基酸。构建了重组表达质粒pET32a/AduMIF-1,经IPTG诱导表达和分离、纯化,获得了重组AduMIF-1,融和蛋白分子质量单位约为33ku。结论本研究从十二指肠钩虫中分离到MIF基因,并成功进行了重组表达、分离与纯化,为进一步研究AduMIF-1的生物学功能奠定了基础。 Objective To clone and express duodenal hookworm macrophage migration inhibitory factor (MIF) AduMIF-1 gene. Methods Specific primers were designed and synthesized. The cDNA of AduMIF-1 gene was amplified by PCR using the cDNA of adult C. elegans. The obtained AduMIF-1 coding sequence was cloned into the prokaryotic expression vector pET32a to construct the recombinant expression plasmid pET32a / AduMIF-1. The recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and induced by IPTG. The recombinant AduMIF-1 was isolated and purified. Results The full-length coding sequence of AduMIF-1 was successfully amplified. The full length reading frame was 360bp and encoded 119 amino acids. The recombinant expression plasmid pET32a / AduMIF-1 was constructed and induced by IPTG. The recombinant AduMIF-1 was obtained, and the molecular mass unit of the fusion protein was about 33ku. Conclusion The MIF gene was isolated from the duodenal hookworm and successfully expressed, isolated and purified, which laid the foundation for the further study on the biological function of AduMIF-1.