Cloning and sequencing of GP50 gene from cysticercus cellulosae

Objective:To set up a way of detecting cysticercosis quickly and accurately by use of recombinant protein,the GP50 gene was amplified from cysticercus cellulosae and cloned into pGEM-T vector.Methods:The specific primers were designed and synthesized.The DNA fragment encoding GP50 gene was amplified by PCR from cDNA of C.cellulosae.The PCR products were cloned into pGEM-T vector and recombinants were screened by PCR,enzyme digestion and sequencing.Results:The size of pGEM-T-GP50 digested with BamH Ⅰ and Xho Ⅰ was identical to the length of the PCR generated products.DNA sequencing indicated that the GP50 gene had an open reading frame of 897 bp,which encodes 298 amino acids with an approximate molecular weight of 33 000 and had high homology with the GP50 gene submitted to GenBank(AY212945).Conclusions:The findings suggested that GP50 gene had been successfully cloned.