Cloning and Nucleotide Sequencing of ScFv Gene for Anti_alpha_toxin of clostridium perfringens Type A

The V H and V L genes were amplified from a hybridoma cell line 1A8 producing mouse McAb against alpha_toxin of Clostridium perfringens type A by RT_PCR.The V H and V L genes were connected through a flexible linker(Gly 4Ser) 3 and the V H_linker_V L(ScFv)fusion gene was cloned into a clone vector pGEM_T.The ScFv gene was sequenced, analyzed by computer. The ScFv gene consists of 729bp encoding 243 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. According Kabat classed method, McAb 1A8 V H gene segment and V L gene segment belong to the mouse lg heavy chain subgroup Ⅱ(A) and κ chain subgroup Ⅵ respectively.