Abstract: Objective To screen the hepatocyte proteins that interact with hepatitis B virus X protein (HBx). Methods The
recombinant plasmid pSos-HBx was constructed by inserting Sos-HBx fragment into the bait vector, and after sequence
verification the plasmid was transformed into competent yeast cells. The expression and self-activation of Sos-HBx protein was
detected in the yeast cells. The hepatocyte proteins interacting with the bait protein was screened with CytoTrap yeast
two-hybrid technique. Results The reconstructed plasmid harboring HBx gene expressed Sos-HBx protein in the yeast cells
without self-activation of the protein. CytoTrap yeast two-hybrid system identified 6 hepatocyte proteins that interacted with
HBx, including fibronectin 1, translationally controlled tumor protein, IQ motif and WD repeats 1, follistatin, orosomucoid 1,
and disulfide isomerase family A member 3. Conclusion Six HBx-binding hepatocyte proteins have been identified using the
CytoTrap yeast two-hybrid system, which provides clues for further investigation of the role of HBx protein in hepatitis and
liver cancer.