2000•Zhongguo shouyi xuebaoRequires access
Cloning and nucleotide sequencing of ScFv gene for anti-alpha-toxin of Clostridium perfringens type A.
Xu Chongbo, Baohua Zhao, Ma Conglin, Feng Shu-zhang, Ping Zhu
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Abstract
The V_(H) and V_(L) genes were amplified from a hybridoma cell line 2E3 producing mouse McAb against alpha-toxin 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 subgroup II (b) and kappa chain subgroup III respectively.