Cloning and Sequencing of V_H and V_L Genes of CD25 McAb

Objective To obtain the VH and VL genes of CD25 McAb and analyze the sequences of them. Methods The VL and VH genes were amplified by RT - PCR from the total RNA of WuTac cell, then cloned into vector PMD18-T and transformed to E. coli JM109 The recombinant plasmids were identified by enzyme digestion , and positive clones were sequenced. Results The result of enzyme digestion was m consis-tent with expected. The cloned VL and VH genes, with the lengths of 324 and 351bp respectively, were belong to the subgroup Ⅳ of light chain and subgroup Ⅲ(C) of heavy chain of murine McAb respectively, and the homology of them to the variable region genes of murine CD25 McAb were above 80% . Conclsion The VH and VL genes of CD25 McAb were directly amplified by RT- PCR in this paper. The study laid a foundation of further development of chimeric and single chain antibodies.