Construction of an expression vector of GPC-3 CTL epitope.
- Author:
Wen-jing WANG
1
;
Cheng-yao LI
;
Ming-song LI
Author Information
- Publication Type:Journal Article
- MeSH: Amino Acid Sequence; Epitopes; chemistry; genetics; immunology; Genetic Vectors; genetics; Glypicans; genetics; immunology; metabolism; Hepatitis B Surface Antigens; genetics; immunology; Plasmids; genetics; metabolism; Polymerase Chain Reaction; T-Lymphocytes, Cytotoxic; immunology; Vaccines, Subunit; chemistry; genetics; immunology
- From: Journal of Southern Medical University 2009;29(8):1548-1550
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the expression vectors of GPC-3 CTL epitope.
METHODSThe 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.
RESULTS AND CONCLUSIONPCR, 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.
