2004•Chinese Journal of VeterinaryRequires access
Cloning and sequencing analysis of ScFv gene for anti-ICAM-1
Yuehong Li, Guoli Zhang, Yue Yu-huan, Qiu Yefeng, Shumin Li, Guangmou Wu, Ping Zhu
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Abstract
The V_H and V_L gene were amplified from a hybridoma cell line F12 producing mouse anti-ICAM-1 McAb by RT-PCR.The V_H and V_L gene were connected through a flexible linker (Gly4Ser)3,and the V_H -link-V_L (ScFv) fusion gene was cloned into a clone vector pMD18-T.The ScFv gene was sequenced,analyzed by computer to be consist of 744 bp encoding 248 amino acid residues.Both V_H and V_L gene were confirmed as functionally rearranged mouse immunoglobulin variable region genes and appeared to be new genes.According to Kabat classed method,McAb F12 V_H gene segment and V_L gene segment belong to the mouse Ig heavy chain subgroup Ⅱ(B) and к chain subgroup Ⅲ,respectively.