Department of Pathogenic Biology and Immunology, Sino-French Hoffmann Institute, School of Basic Medical Science, Guangzhou Medical University, Xinzao Town, Panyu, Guangzhou 511436, Guangdong, China
Department of Scientific Research and Education, Yuebei People’s Hospital, Shaoguan 512026, Guangdong, China.
Department of Clinical Laboratory Sciences, Renmin Hospital, Hubei University of Medicine 442000, Hubei, China
Department of gynaecology, Yuebei People’s Hospital, Shaoguan 512026, Guangdong, China.
Background: Recently, the incidence of allergic diseases has been on the rise; Dust mite is the major indoor allergen which needs a special consideration.
Objectives: This study was carried out to identify and investigate the molecular properties of a new allergen named Hsp60 and to afford a foundation for future research of the allergic diseases caused by Dermatophagoides farinae.
Materials and Methods: Using polymerase chain reaction (PCR) with degenerate primer, the cDNA of Dermatophagoides farinae Hsp60 was amplified and sequenced. Next, the cDNA fragment was cloned into the prokaryotic expression vector pET-32a for the expression of the Hsp60. Then, it was further characterized by Elisa and Western Blot analysis.
Results: The partial cDNA sequence of the Dermatophagoides farinae Hsp60 was determined, and the recombinant Hsp60 was successfully expressed in Escherichia coli BL21. ELISA and Western blot analysis showed that the recombinant protein could be specifically recognized by SIgE from sera of the Dermatophagoides farina-allergic patients.
Conclusions: Our group has, for the first time, demonstrated the fact that there is an Hsp60 family of Dermatophagoides farinae and analyzed the allergenicity of the Hsp60 with immunological method. These results provide a foundation for further allergological research of the Dermatophagoides farinae Hsp60.