Abstract: The purpose of this study was to construct recombinant adenovirus vector of co-expression MRP and EF genes from Streptococcus suis type 2（SS2）. The specific primers were designed by the sequence of MRP and EF genes. The MRP（1 801-2 513）and EF（1 783-2 563）gene fragment were amplified from genomic DNA of SS2 by PCR technique. PCR products were cloned in pMD18-T vector. Then MRP gene, IRES element and EF gene linked into the adenovirus shuttle plasmid（pShuttle-CMV）to construct recombinant shuttle plasmid（pShuttle-CMV-MRP-IRES-EF）. The recombinant shuttle plasmid was lineared with PmeⅠand transformed into BJ5183-AD-1 competent cells to construct homogeneous recombinant adenovirus plasmid（rAdeno-CMV-MRP-IRES-EF）. Then the recombinant adenovirus plasmid was linearized with PacⅠand transfected into AD-293 cells. Finally, virus liquids of cell pack were identified by PCR. The results showed that MRP and EF genes were verified by gene sequencing. Both the recombinant shuttle plasmid and recombinant adenovirus vector plasmid were verified by enzyme digestion. Cytopathic effect（CPE）was observed after transfection linear rAdeno-CMV-MRP-EF 6 days. MRP and EF gene were also detected in the virus liquid by PCR.

Key words: Streptococcus suis type 2, Muramidase-released protein, Extracellular factor, Gene co-expression, Recombinant adeno-virus vector