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Updated: May 28, 2026
Removal and Replacement of Endogenous Ligands from Lipid-Bound Proteins and Allergens
Published on: February 24, 2021
[Cloning, expression and purification of allergen parvalbumin from Aristichthys nobilis and its allergic activity]
Haiyan Wang1, Li Li, Zhigang Liu
1Institute of Allergy and Immunology, Shenzhen University, Shenzhen 518060, China. haitun107@gmail.com
Objective:
To clone, express and identify the parvalbumin gene from Aristichthys nobilis, and investigate its allergenicity.
Methods:
The parvalbumin gene was amplified by RT-PCR and cloned into PMD18-T for sequencing and analysis. Then the target gene was subcloned into pET-28a (+) for expression in E. coli BL21 (DE3) by IPTG induction. The recombinant protein was purified by metal (Ni2+) chelating affinity chromatography. Its allergenicity was examined by Western-blotting assay.
Results:
The length of gene (Accession No. FJ013047) was 330 bp, coding 109 amino acids. The E. coli strain could express a recombinant protein with a molecular weight of 11 537 Da. The recombinant allergen was identified as its affinity to specific IgE antibodies from the allergic patient sera by Western-blotting.
Conclusion:
The parvalbumin gene from Aristichthys nobilis is successfully cloned and expressed in this study, and the recombinant protein possesses good IgE-binding capacity.
