σC gene expresson of muscovy duck reovirus MW9710 strain in Pichia pastoris

The σC gene of DRVMW9710 was inserted into pPIC9K to construct a recombinant secretory vector,pPIC9K-σC.Then,pPIC9K-σC was transformed into E. coli DH5α.The recombinant vector was identified by PCR,enzymatic analysis and sequencing.The positive plasmid was linearized by Sac I and transformed into GS115 by electroporation.The insert was integrated into genomic DNA of Pichia pastoris by means of homologous recombination.The recombinants with multiple integrated copies of σC were screened using gradient concentration G418,and inducted with methanol.The results of SDS-PAGE and Western-blot showed that the highly expressed protein had an immunoreaction property.