CLONING AND SEQUENCING ANALYSIS OF VARIABLE REGION GENES OF ANTI-DIFLOXACIN McAb

The variable region genes of heavy chain(VH) and light chain(VL) of anti-difloxacin monoclonal antibody(McAb) were amplified by RT-PCR from total RNA extracted from X1 hybridoma cell.PCR products were cloned into pUC18 vector and sequenced by Sanger's dideoxymediated chain-termination method.The results showed that VH gene consisted of 363bp encoding 121 amino acid residues;VL gene consisted of 348bp encoding 116 amine acid residues,which contain four FRs and three CDRs,respectively.Compared with mouse Ig database,the VH and VL regions are in accord with the characterization of DNA sequence present in the mouse Ig region.The successful clone of VH and VL genes of X1 McAb made it a solid foundation to the construction of engineering antibody against difloxacin.