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Updated: May 28, 2026
Characterization of Glycoproteins with the Immunoglobulin Fold by X-Ray Crystallography and Biophysical Techniques
Published on: July 5, 2018
[Prokaryotic expression, purification and antigenicity identification of human GRP78 protein]
Hui-juan Song1, Hong-dan Li, Jia Wei
1Liaoning Medical University, Jinzhou, China. shj-shj@tom.com
Aim:
To construct a recombinant plasmid pGEX-4T-1-GRP78, express it and purify human glucose regulated protein 78 (GRP78).
Methods:
GRP78 gene was amplified by PCR from the recombinant vector constructed in our laboratory. The PCR product was inserted into pGEX-4T-1 prokaryotic expression vector to generate pGEX-4T-1-GRP78. The pGEX-4T-1-GRP78 was then transformed into BL21 and GRP78 was expressed on induction of IPTG. After purification, GRP78 was released by thrombin cleavage, and its antigenicity was identified by ELISA.
Results:
The GRP78 prokaryotic expression vector was successfully constructed as confirmed by enzyme digestion and DNA sequencing. ELISA demonstrated the antigenicity of the purified GRP78 protein.
Conclusion:
The recombinant prokaryotic expression vector pGEX-4T-1-GRP78 has been constructed successfully. The purified GRP78 has been obtained with good antigenicity, which will be used for GRP78-based serum diagnosis of hepatocellular carcinoma.
