Objective　To construct a lentivirus vector for RNA interference (RNAi) of BC047440 gene

and detect the silence effect of the vector in human hepatocellular liver carcinoma cell line (HepG2).  　Methods　The targeting sequence of BC047440 gene which can be effectively silenced in RNA inference was confirmed in our previous study. The cDNA encoding for sense and

antisense Oligo DNA fragments of the targeting sequence was designed

synthesized and cloned into the pFU-GW-iRNA vector which had been double digested by the HpaⅠ and XhoⅠ

and vector then the results  confirmed by PCR and DNA sequencing. 293T cells were cotransfected with lentiviral vector pFU-GW-shBC047440

pHelper 1.0 and pHelper 2.0 by Lipofectamine 2000

to produce lentiviral vector which can infect other cell lines. The virus titer and the fittest multiplicity of infection (MOI) in HepG2 were measured by the expression level of green fluorescent protein (GFP). The infected HepG2 was divided into BC047440-shRNA group

control-shRNA group and HepG2 group. The changes of BC047440 mRNA and protein level were detected by RT-PCR and Western blot analysis respectively.  　Results　The data of PCR demonstrated that the lentivirus RNAi vector of BC047440 was constructed successfully. The titer of virus was 5×10

8

TU/ml

and the fittest MOI in HepG2 was 20. BC047440 and BC047440 mRNA expression of BC047440-shRNA group was obvious lower than the control-shRNA group and HepG2 group

but there was no visible different between the control-shRNA group and HepG2 group.  　Conclusion　The lentivirus RNAi vector of BC047440 is constructed successfully. After this lentivirus vector infects HepG2 cells

the target of silencing the expression on BC047440 is achieved.