Cloning and sequcllcing of the VH/VL genes of antihuman γ-seminoprotein McAb

Aim:To clone the variable region gene of murineMcAb against γ-seminoprotein, for constructing the singlechain variable fragment. Methods:Total RNA was extractedfrom the hybridoma cell line E4B7, which secretes McAb against γ-seminoprotein, and subjected to reverse transcription. The VH and VL genes were amplified by PCR, clonedinto plasmid vector and analysed using auto DNA sequencer. Results: E4B7 VH consists of 358 hp encoding 119amino acid residues, E4B7 VL contains 326 hp encoding 108amino acid residues. The deduced amino acid sequences ofE4B7 VH/VL were in agreement with the characterization ofthe amino acid present in the mouse Ig variable region. Conclusion: The cloned sequences were both confirmed as newgenes by comparing with all known genes in GenBank in Internet. The sequences exhibite significant homology to previously reported antibody variable region genes.