[Construction of an expression vector of GPC-3 CTL epitope].
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.
Wang
Wen-jing
WJ
School of Biotechnology, Southern Medical University, Guangzhou 510515, China. wenjingking@163.com
Li
Cheng-yao
CY
Li
Ming-song
MS
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
D000939:Epitopes; D053673:Glypicans; D006514:Hepatitis B Surface Antigens; D022223:Vaccines, Subunit