title: [Construction of an expression vector of GPC-3 CTL epitope].
abstract: OBJECTIVE
  To construct the expression vectors of GPC-3 CTL epitope.
  
  
  METHODS
  The HBsAg gene with three different EYILSLEEL (EYI) sites was named EYI1, and another with one EYI replacing CTL epitope FLG or SIL of pcHBsAg were named EYI2 and EYI3, respectively. All the three DNAs were amplified by SEOing PCR from pcHBsAg plasmid and linked into pBSSK+ vector to construct Pbssk/EYI1, pBSSK/EYI2, and pBSSK/EYI3. The three plasmid were identified by PCR, double digestion and sequencing, and the fragments with EYI1-3 were obtained by double digestion and then inserted into pcDNA3.1+ vector.
  
  
  CONCLUSIONS
  PCR, enzyme digestion and sequence analysis confirmed successful construction of the eukaryotic expression vectors pcDNA-EYI1/HBsAg, pcDNA-EYI2/HBsAg, pcDNA-EYI3/HBsAg, which facilitate further studies of the GPC3-HBsAg multiple peptides vaccine for HBV infection.
authors.affiliation: School of Biotechnology, Southern Medical University, Guangzhou 510515, China. wenjingking@163.com
authors.forename: Wen-jing
authors.initials: WJ
authors.lastname: Wang
authors.forename: Cheng-yao
authors.initials: CY
authors.lastname: Li
authors.forename: Ming-song
authors.initials: MS
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000939:Epitopes / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D005822:Genetic Vectors / Q000235:genetics*; D053673:Glypicans / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D022223:Vaccines, Subunit / Q000737:chemistry / Q000235:genetics / Q000276:immunology
chemical_list: D000939:Epitopes; D053673:Glypicans; D006514:Hepatitis B Surface Antigens; D022223:Vaccines, Subunit