Construction and expression of HBV preS1 gene in yeast

Objective: To construct the yeast expression vector of HBV preS1 gene for investigating the potential role of preS1 protein in mediating the attachment of HBV particles to human hepatocytes. Methods: PCR was performed to amplify HBV preS1 gene from the plasmid PCP10/HBV ayw subtype containing the whole fragment of HBV, and the PCR product was cloned into pGEM-T vector and then named T-preS1. HBV preS1 gene was cut from T-preS1 by Nco I and Mlu I, and cloned into yeast expression plasmid pSos; the reconstructed plasmid was tested by auto-sequencing assay and named pSos-preS1. pSos-preS1 was transformed into yeast cell cdc25 (a) by LiAc mediated transformation, and the yeast protein was isolated and analyzed by Western blot. Results: The yeast expression vector of HBV preS1 gene was constructed successfully, and the presence of HBV preS1 protein in yeast cells was confirmed by Western blot analysis. Conclusion: Reconstruction of pSos-preS1 has laid a foundation for better understanding of the mechanism of HBV preS1 protein in viral endocytosis and is helpful in seeking the preS1 related protein that is supposed to interact with preS1 protein in vitro by sos-recruitment system.