Validation On The Interaction Between Jumping Translocation Breakpoint Protein And Envelope Protein Of Hepatitis B Virus

OBJECTIVE:The present research aims at validating the interaction between jumping translocation breakpoint (JTB) protein and envelope protein of hepatitis B virus (HBsAg) by using mammalian two-hybrid, co-immunoprecipitation, GST pull-down.METHODS: Mammalian Two-Hybrid Assay Kit produced by Stratagene Co. Ltd was adopted in the present research. The plasmid pCMV-BD-JTB, pCMV-AD-S and report plasmid pFR-Luc were co-transferred into 293FT cell line to evaluate the interaction between JTB protein and HBsAg. BD-JTB and AD-S infusion proteins were collected for bidirectional co-immunoprecipitation. Further validation was performed by using GST pull-down.RESULTS:According to the result of mammalian two-hybrid, expression of luciferase was higher in co-transfection of HBsAg and JTB protein compared with positive control. The results co-immunoprecipitation, GST pull-down indicated that there is an interaction between JTB protein and HBsAg. And the interacting site may locate at the domain of main HBsAg.CONCLUSION:There is an interaction between JTB protein and HBsAg. The interacting site may locate at the domain of main HBsAg.