Screening of gene encoding of hepatic proteins interacting with pre-S1 transactivated protein 3 via yeast two-hybrid

Objective To screen proteins in leukocytes interacting with hepatitis B virus(HBV) pre-S1 transactivated protein 3(PS1TP3)by yeast-two hybrid.Methods The DNA fragment of PS1TP3 was amplified by polymerase chain reaction.Bait plasmid pGBKT7-PS1TP3 was constructed and transform it to yeast cells AH109.Then mated it with yeast cells Y187 containing epatic cDNA library plasmid pACT2.The obtained diploid yeast cells were plated on synthetic dropout nutrient medium SD/-Trp-Leu-His-Ade containing X-α-Gal for selection twice.The plasmids of positive colonies were transformed to E.coli and digested by Bgl II enzyme,then sequenced and analyzed by bioinformatics.Results Thirty proteins binding to PS1TP3 were screened,including ubiquitin-conjugating enzyme E2,mitogen activated protein kinase,beta-2-microglobulin,phosphatase and tensin etc gene whose functions had been known and 4 unknown function series and one splice new gene.Conclusion Conjugated protein of PS1TP3 are successfully cloned and the results bring some new clues for studying the molecular biological functions of PS1TP3 and pathogenic mechanism of HBV.