Expression of Two Gonadotropin β-subunits of Acipenser schrenkii in E.coli

Using the genetic engineering method,two gonadotropin β-subunit of Acipenser schrenkii were exprssed in E.coli.Two cDNA Sequenes of gonadotropin(GTH) β-subunits mature peptide cDNA sequences of Amur sturgeon were inserted into pET-22b(+) vector,using NdeⅠ and XhoⅠ restriction site.The recombinant plasmids of two GTH β-subunits were constructed as C-terminal histidine-tagged fusion protein.Transformants harboring expression vectors were induced in E.coli strain BL21(DE3),the recombinant GtHⅠ-β and GTHⅡ-β were about 14 000 and 15 000 respectively.The results of SDS-PAGE and western-blotting with the McAb of 6-histidine showed that the recombinant protein had correct molecular weight,just as expected.They also showed high immunological activities through western-blotting with polyclonal antibody of purified sturgeon GTH. GtHⅠβ recombinant protein had obvious expression in 2 hours.The quantity of induced protein had no significant increase after 6 hours.The combinated protein will contribute to develop a RIA method for detection of Acipenser.