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 / 中华微生物学和免疫学杂志
Chinese Journal of Microbiology and Immunology
; (12): 340-343, 2010.
Article
in Zh
| WPRIM
| ID: wpr-379884
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WPRO
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.
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WPRIM
Language:
Zh
Journal:
Chinese Journal of Microbiology and Immunology
Year:
2010
Type:
Article