Aim: To reveal the influence of HCV-NS5A on PI3K, we probe into the relationship between NS5A and PI3K in vitro.
Methods: Full length NS5A gene of HCV was amplified by PCR, using the plasmid containing HCV full-length open reading frame (ORF) as template, and cloned into the eukaryotic expressing plasmid pcDNA3.0(-) by DNA recombination technique. The recombinant vector was identified by digestion with restriction enzymes and polymerase chain reaction and by directly sequencing. Then both the recombinant vector pcDNA3.0(-)-NS5A and the control vector pcDNA3.0(-) were transfected HepG2 cell using Lipofectamin2000.
Results: Expressing NS5A was proven by RT-PCR and Western blot analysis in transfected HepG2 cell. Further more we examined the PI3K protein expressed in the HepG2 cell expressing recombinant NS5A.
Conclusion: NS5A can activate PI3K in vitro and its signal cascade pathway.