Cloning and expression method for Ani s4 antigen gene of Anisakis simplex

The invention especially relates to a cloning and expression method for the Ani s4 antigen gene of Anisakis simplex, which belongs to field of biotechnology. According to the method, total RNA of third-stage larvae of Anisakis simplex is used as a template; a specific fragment with a size of 500bp is obtained through amplification by using a RT-PCR method; the amplification product--specific fragment is cloned into the pMD18-T transformation competent bacterium DH5-alpha; enzyme digestion and identification are carried out so as to obtain positive recombinant plasmid; the recombinant plasmid and an expression vector pET-32a are respectively subjected to BamHI and HindIII double enzyme digestion so as to construct a recombinant expression vector pET-32a-Ani-S4; the recombinant expression vector pET-32a-Ani-S4 is transformed into Escherichia coli BL21; plasmid is extracted and undergoes enzyme digestion and PCR identification, and after it is proved the plasmid is correct, IPTG induced expression is carried out; SDS-PAGE and Western-blot test of an expressed product show that fusion protein has molecular weight of about 25 kDa, so the Ani s4 gene is successfully expressed in Escherichia coli. The His-tag of the constructed expression protein brings great convenience to subsequent purification. Successful expression of the ES antigen protein lays a foundation for subsequent research on immunological detection methods for Anisakis simplex.