Abstract: Objective:To construct the recombinant plasmid vector of pcDNA3.1-RPS27a,and investigate the localization and expression of RPS27a protein in HepG2 and SMMC7721 cell lines.Methods:The RPS27a gene was synthesed and cloned into the pcDNA3.1 vector.After the pcDNA3.1 plasmid was transfected into E.coli DH5α,and the positive plasmid was extracted and sequenced.The reconstructed pcDNA3.1-RPS27a plasmid was transfected into the HepG2 and SMMC7721 cell lines.The expression and localization of RPS27a in eukaryotic cells were detected by laser confocal microscopy.Results:The recombinant plasmid pcDNA3.1-RPS27a was successfully constructed.The laser confocal microscopy showed that the RPS27a mainly expressed in the cytoplasm of HepG2 and SMMC7721 cell lines.Conclusions:The recombinant plasmid pcDNA3.1-RPS27a is successfully constructed,and the RPS27a protein mainly expresses in the cytoplasm of HepG2 and SMMC7721 cell lines.