Results
Expression,
purification and identification of SADS-CoV S protein
To express the S protein of SADS-CoV in vitro , eukaryotic
expression of the synthesized S gene was done using the vector
pCMV with an Fc tag (Fig 1), then the cell lysates infected by rB or wB
was detected by Western Blot with anti-huamn Fc PcAb and porcine
anti-SADS-CoV positive or negative serum, respectively. As shown in Fig
2A, a specific band could be detected by Western Blot, indicating that
S-Fc protein have been expressed and have good antigenicity. Since the
S-Fc protein was confirmed successfully expressed in HEK 293F cells, the
S-Fc protein was purified with protein G Resin. As shown in Fig 2B, more
95% purity protein was examined by SDS-PAGE, indicating that purified
S-Fc protein can be used to develop ELISA.