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Cloning and characterization of tropomyosin, an allergen of periplaneta fuliginosa

Title
Cloning and characterization of tropomyosin, an allergen of periplaneta fuliginosa
Other Titles
먹바퀴(Periplaneta fuliginosa) 알레르겐, 트로포마이오신의 클로닝 및 특성규명.
Issue Date
2003
Publisher
Graduate School, Yonsei University
Description
Dept. of Medicine/박사
Abstract
[한글] 주가성 절지동물은 알레르기 질환의 가장 중요한 원인이다. 한국에는 네 종의 주가성 바퀴가 보고되어 있으나, 현재까지의 연구는 독일바퀴와 이질바퀴에 국한되어 있다. 독일바퀴 및 큰다리먼지진드기에 감작된 환자 혈청을 이용하여 억제효소면역흡착법과 면역이적법을 수행하여 먹바퀴와의 교차반응을 조사하였다. 모든 30명의 환자혈청이 먹바퀴와 반응하였으며, 먹바퀴의 IgE 반응이 독일바퀴에 의해 79.4%, 큰다리먼지진드기에의해 63.3% 억제되었다. 교차반응이 강하여 pan-allergen으로 알려진 트로포마이오신 cDNA를 RT-PCR로 클로닝하고, pET-28b 발현시스템을 이용하여 재조합 단백질을 생산하였다. 연역한 아미노산서열이 이질바퀴 트로포마이오신과 일치 하였다 (뉴클레오타이드 서열은 98% 일치). 재조합단백질은 26개의 환자혈청 중에서 7개의 환자혈청과 특이적으로 반응하였으며, 10 ㎍/ml의 농도에서 조항원의 IgE 반응을 37.7% 억제하였다.먹바퀴는 독일바퀴 및 큰다리먼지진드기와 강한 교차반응을 하는 알레르겐을 가지는 것을 알 수 있었으며, 교차반응성과 알레르기발현성을 보다 잘 이해하기 위해서는 순수한 트로포마이오신을 분리하여 자세한 연구를 수행할 필요가 있다. 재조합트로포마이오신은 앞으로의 연구와 임상응용에 유용할 것으로 사료된다.
[영문] Household arthropods are one of the most common causes of allergic diseases. Four species of cockroaches are found to reside in Korean homes, but published work almost deals exclusively with the German and American cockroaches. This study was undertaken to investigate the cross-reactive allergenic components of dusky brown cockroach, Periplaneta fuliginosa. ELISA inhibition and immunoblot analyses for P. fuliginosa were performed with Blattella germanica and Dermatophagoides farinae allergic sera to investigate the cross-reactive allergens. All 30 sera tested showed that P. fuliginosa-specific IgE, and IgE-binding reactivity of P. fuliginosa were inhibited up to 79.4% by B. germanica and up to 63.3% by D. farinae. cDNA encoding tropomyosin, which is a well known cross-reactive pan-allergen, was cloned by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), and its recombinant protein was produced using a pET-28b expression system. The deduced amino acid sequence of cloned cDNA was identical with that of P. americana tropomyosin (with 98% nucleotide sequence identity). Seven of 26 (26.9%) allergic sera had specific IgE for the recombinant protein, and the maximum inhibition of P. fuliginosa specific IgE achieved with recombinant tropomyosin was 37.7%, at an inhibitor concentration of 10 ㎍/ml. P. fuliginosa appears to possess allergens, which are highly cross-reactive allergens with allergens of B. germanica and D. farinae. More detailed studies using native tropomyosin are needed to further understand its cross-reactivity and allergenicity. Recombinant tropomyosin could be useful for further studies and clinical applications.
URI

http://ir.ymlib.yonsei.ac.kr/handle/22282913/128358
Appears in Collections:
2. 학위논문 > 1. College of Medicine (의과대학) > 박사
Yonsei Authors
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