Ultraviolet cross-linking assay in screening of cellular proteins binding to Hepatitis C virus RNA

  Objective   To select a better method for screening of cellular proteins binding to Hepatitis C virus(HCV)RNA.

  Methods   DIG-labeled RNA(DIG-RNA)transcripts were prepared by in vitro transcription.Cytoplasmic extracts were prepared from human hepatoma cells(HepG2).Ultraviolet(UV)cross-linking assay and electrophoretic mobility shift assay(EMSA)were used to screen the cellular proteins that would bind to HCV RNA.

  Results   Both UV cros-slinking assay and EMSA detected that there were proteins binding to HCV RNA.The signals of DIG-RNA detected by EMSA were dispersing and unclear.The results of UV cross-linking assay were clear RNA-binding protein bands.

  Conclusion   UV cross-linking assay is a preferred method for screening of cellular proteins binding to Hepatitis C virus RNA and other RNA molecules.