Construction of SCFV gene against surface memberane antigen of Eimeria acervulina sporozoite

To construct gene ScFv (single chain variable fragment) directed against Eimeria acervulina sporozoite surface memberane antigen, the V-H and V-I. gene were amplified, By means of RT-PCR technique, from a hybirdoma cell line Easp - 3G3 producing murine McAb against Eimeria acervulina sporozoite. To obtain a functional single-chain antibody (scFv), a linker region consisting of (Gly4Ser) 3 was introduced by gene splicing by overlap extension (SOE) and the V-H-linker-V-L (ScFv) fusion gene were cloned into a clone vector pMD-18T and sequenced. The ScFv gene was sequenced, analyzed by computer to be consisted of 744 bp encoding 248 amino acid residues. Both V-H and V-L genes were confirmed as functionally rearranged mouse immunoglobulin variable region genes and appeared to be new genes. The single chain variable Fv of a monoclonal antibody against Eimeria acervulina sporozoite was constructed successfully. This ScFv might be as a new tool in researching molecular nosogenesis of coccidiosis.