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 2E3 producing mouse McAb against alpha txoin of Clostridium perfringens by RT PCR. The V H and V L genes were connected through a flexible linker (Gly4Ser)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 to be consist of 726 bp encoding 242 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 2E3 V H gene segment and V L gene segment belong to the mouse Ig heavy chain subgroupⅡ(b) and κ chain subgroup Ⅲ respectively.