Construction and appreciation of eukaryotic expression vector of HBV prec/c shRNA

Objective To investigate new measures to prevent and cure the infection of hepatitis B virus and construct eukaryotic expression vectors shRNA aiming directly at the pre-core gene sequence of hepatitis B virus through observing its effect on HBeAg expression in HepG22.15 cells.Method The shRNA express vectors of psiHBV1,psiHBV2 and psiHBVc aiming directly at the pre-core gene sequence of hepatitis B virus were constructed.After psiHBV with lentiviral vector auxilary system was transfected into 293T cells,the lentiviral particles were made up and infected HepG22.15 cells.The transcription of prec/c Mrna and the expression of HBeAg in cell supernatant and cell lysate were detected by the methods of RT-PCR and MEIA respectively.Result The identification of shRNA by enzyme cutting and sequencing was coincided with the anticipation.After the lentiviral particles were assembled and HepG2 2.15 cells infected the transcription of prec/c mRNA and the expression of HBeAg decreased greatly.Compared with control group the inhibition of lentiviral particles to expression of HBeAg showed significant deviation(P0.01) Conclusion The lentiviral vectors psiHBV1,psiHBV2 and psiHBVc were constructed succesfully.RNAi mediated by lentivirus can control the expression of HBV.It provides a basis for the application of RNA interference in treatment of hepatitis B.