Fusion expression of the extracellular carbohydrate recognition domain of mouse Dectin-1 and its recognition of β-glucans in the cell wall of Candida albicans

Abstract

Objective To clone, express and purify the extracellular carbohydrate recognition do-main(CRD) of Dectin-1 in mouse peritoneal macrophages and to further investigate its ability to recognize and bind to β-glucans. Methods The Dectin-1 CRD gone was amplified by RT-PCR from RNA of mouse peritoneal macrophages and cloned into prokaryotic expression vector pET28a (+), the constructed pET-CRD recombinant plasmid was transformed into Escherichia coli BL21 (DE3) and the fusion protein was in-duced to express. After affinity purification and renaturation, the fusion protein was incubated with Candida albicans yeast and its ability to recognize and bind to β-glucans in the cell wall of fungi. Results The fu-sion protein could recognize β-glucans in the fungal cell wall. Conclusion The recombinant expression plasmid pET28a-CRD was successfully constructed and the fusion protein was induced. The fusion protein is able to recognize and bind to β-glucans in the fungal cell wall, thus laying a good foundation for fungal de-tection and the exploration of the biological role of β-glucans. Key words: Dectin-1; β-glucans; Fusion expression; Candida albicans