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Updated: Jun 20, 2026
A High Yield and Cost-efficient Expression System of Human Granzymes in Mammalian Cells
Published on: June 10, 2015
[Construction of an expression vector of GPC-3 CTL epitope]
Wen-jing Wang1, Cheng-yao Li, Ming-song Li
1School of Biotechnology, Southern Medical University, Guangzhou 510515, China. wenjingking@163.com
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.
Results And Conclusion:
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.
