| Goose paramyxovirus disease is an acute contagious disease caused by GPMV,a member of Rubulavirus family.GPMV has been found in many areas since it was reported by Wang Yongkun of Yangzhou university in 1997,which was characterized by the lesion of digestive tract with high mortality and occurrence,and has caused a large loss in economy.The primary study show that there is a difference in pathogenicity between GPMV and traditional NDV,and the common NDV vaccine does not provide an effective protection against disease.So it is of great interest to study the pathogen from the molecular biology. In this study,according to the complete genome sequence of GPMV SF02 isolate and other NDV strain published in GenBank,two pair of primers were desighed to amplify M gene by RT-nested PCR.The specific DNA products were cloned into pMD18-T vector,and transformed into competent cell TG1,and the positive recombinant plamids were screend by AMP/IPTG/X-gal,and endonuclease digesting and PCR identification.The sequence analysis showed that M gene is 1095 bp in length,encoding 365 amino acid residues;the homology of nucleotide between JS/1/97/Go strain and other strains varies from 84.8%～98.9%. According to date specific major antigen site of M is in 5′extremity 444 bp in length,another pair of primers was designed to amplify M gene of major antigen site (1bp-444bp). The major antigen site of M gene was into expression plasmid pGEX-6P-1.The recombinant plasmid pGEX-6P-M1 was transformed into E.coli BL21(DE3)PlysS and induced with IPTG.A fusion protein as we expected was found. The expressed products were tested by western blot.The results showed that GST-M1 fusion protein has a positive reaction. This study is first time in amplify complete JS/1/97Go strain /M gene and get express of major antigen site of M,it provides the basis evidence and material basis for GPMV identification,molecular epdiemiology investigation research of diagnostic reagent.
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