Construction and identification of prokaryotic expression plasmid pCold-TF-Der f 1 for the dust mite allergen Der f 1

Objective To construct prokaryotic expression plasmid pCold-TF-Der f 1 for the dust mite allergen Der f 1. Methods The Der f 1 gene in full length was amplified from pET-28a(+)-Der f 1 plasmid and cloned to the vector pCold-TF-DNA, with the recombinants then transferred into Escherichia coli BL21. The genetically engineered bacteria with pCold-TF-Der f 1 plasmids were induced by IPTG and the expressed products were analyzed by SDS-PAGE and Western blotting. Results The Der f 1 gene was acquired by PCR, with pCold-TF-Der f 1 successfully constructed, and expressed in E. coli. which was indicated SDS-PAGE and western blotting. Conclusion The prokaryotic expression plasmid pCold-TF-Der f 1, which contain Der f 1 gene fragment of Dermatophagoides farinae, can be successfully constructed with its expression in E. coli BL21 accomplished.