Cloning and Expression of the Amino Structure Domain Gene of Sialoadhesin from Swine and Preparation of Polyclonal Antibody against it

The pig sialoadhesin near the amino structure domain cDNA sequence which was cloned from the 90-day age lung health piglets macrophages with RT-PCR was subcloned into a prokaryotic expression vector pET-32a,to construct a recombinant expression plasmid pET-Sn150 which was successfully used to express a gamma recombinant protein subunits whose molecular weight was about 43 ku.By using the recombinant protein pET-Sn150 to immune the mouse,the mice sialoadhesin resistance gamma-irradiation His recombinant protein polyclonal antibody which could be tested by Western blotting and indirect ELISA could be achieved.And the result of indirect ELISA showed that the titer of the polyclonal antibody was 1∶12800.The specific binding of the mice sialoadhesin resistance gamma-irradiation His recombinant protein polyclonal antibody with the restructuring γ and unit protein revealed by the result of Western blotting proved that the recombinant protein had good immunogenicity,and that layed the foundation for a further study of its construction and function as the pig sialoadhesin near the amino structure domain gene was successfully cloned and expressed.