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한국산 집먼지진드기 주알레르겐의 분리 및 합성

Other Titles
 Purification of the major allergens from Korean Dermatophagoides pteronyssinus and production of the recombinant antigens 
Authors
 김경섭  ;  박상욱  ;  박중원  ;  홍천수  ;  오상환 
Citation
 Journal of Asthma, Allergy and Clinical Immunology (천식 및 알레르기), Vol.19(1) : 91-102, 1999 
Journal Title
 Journal of Asthma, Allergy and Clinical Immunology (천식 및 알레르기) 
ISSN
 1226-8739 
Issue Date
1999
Keywords
house dust mite ; Dermatophagoides pteronyssinus ; recombinant protein allergen ; purification
Abstract
Background: Purified major allergens of house dust mite are essential for evaluation of the allergic mechanism in molecular basis and development of new modalities of immunemodulation. Objective: In this study, we aimed to purif group 1 and group 2 allergens from Dermatophagoides pteronyssinus (Dp). In addition, cDNAs corresponding to Der pI and II in Korean Dp were isolated and recombinant Der p1 and Der pII were synthesized. Materials and method: Der pI allergen was purified by ammonium sulfate precipitation, anion -exchange column chromatography, and gel filtrat,ion chromatography. Der pII allergen was purified by anion exchange chromatography, gel filtration chromatography, and a preparative isoelectric focusing method. Results: Eight hundred ug of Der pI and 50 ug of Der pII were obtained from 100 g of culture medium and 1 g of mite bodies, respectively. The purities of these allergens were confirmed by SDS PAGE and the strong reactivity to the patient sera was identified. In order to produce a recombinant allergens, poly(A) RNA from house dust mites were isolated and used for cDNA synthesis by RT PCR. The cDNA was inserted into prokaryotic expression vector and the vectors were transformed into E. coli. A little amount of recombinant Der pI protein was produced due to the low solubility, and 1.2 mg of recombinant Der pII was produced from 1 L of E. coli culture medium. The antigenicity of Der pI was relatively weak, however, Der pII showed a strong antigenicity. Amino acid sequence of the amplified cDNA deduced from DNA sequences of Der pII showed 6 different variants. The variation of amino acid sequences suggests the possibility of high incidence of mutation of Der pII protein. Conclusion: A simplified method for the purification of Der pI and Der pII was developed. Recombinant allergens will be useful for the diagnosis and treatment of allergy with lower costs.
Files in This Item:
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Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Biochemistry and Molecular Biology (생화학-분자생물학교실) > 1. Journal Papers
Yonsei Authors
Kim, Kyung Sup(김경섭) ORCID logo https://orcid.org/0000-0001-8483-8537
Park, Sahng Wook(박상욱) ORCID logo https://orcid.org/0000-0002-9594-7074
Hong, Chein Soo(홍천수)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/172581
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