Allergenicity of recombinant Bla g 7, German cockroach tropomyosin
Allergenicity of recombinant Bla g 7, German cockroach tropomyosin
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DOI：
10.1034/j.1398-9995.2003.00167.x
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发表时间：
2003-10-01
期刊：
影响因子：
12.4
通讯作者：
Yong, TS
中科院分区：
文献类型：
--
作者：
Jeong, KY;Lee, J;Yong, TS
背景：蟑螂侵扰可能使易感个体敏感并引起过敏反应。无脊椎动物原肌球蛋白是过敏的常见原因，在自然界中具有高度的交叉反应。本研究旨在制备重组德国蟑螂原肌球蛋白，并对其致敏性进行研究。方法：采用逆转录聚合酶链式反应（RT-PCR）克隆德国蜚蠊原肌球蛋白（Bla g 7）。克隆的cDNA在大肠杆菌中过表达，用Ni-nitrilotriacetic （NTA）酸性树脂亲和层析纯化。采用酶联免疫吸附试验（ELISA）检测重组原肌球蛋白的致敏性。结果：克隆的bla7与其他蟑螂原肌球蛋白氨基酸序列同源性高达91%。ELISA结果显示，重组bla7对德国蜚蠊过敏血清的致敏率为16.2%。重组原肌球蛋白能抑制32.4%的特异性IgE与蟑螂提取物的结合。结论：原肌球蛋白是蟑螂提取物中一种次要的过敏原。希望重组原肌球蛋白能够为进一步的研究和临床应用提供有益的指导。
Background: Cockroach infestation may sensitize and elicit allergic responses to genetically predisposed individuals. Invertebrate tropomyosins are a frequent cause of allergy and highly cross-reactive in nature. In this study, we aimed to produce recombinant German cockroach tropomyosin and investigate its allergenicity.Methods: German cockroach tropomyosin (Bla g 7) was cloned by reverse transcriptase polymerase chain reaction (RT-PCR). The cloned cDNA was overexpressed in Escherichia coli and purified by affinity chromatography using Ni-nitrilotriacetic (NTA) acid resin. The allergenicity of the recombinant tropomyosin was examined by enzyme-linked immunosorbent assay ( ELISA).Results: The cloned Bla g 7 shared up to 91% amino acid sequence identity with other cockroach tropomyosins. ELISA showed a recombinant Bla g 7 sensitization rate of 16.2% to German cockroach allergic sera. Recombinant tropomyosin was able to inhibit 32.4% of the specific IgE binding to cockroach extract.Conclusions: Tropomyosin represents a minor allergen in cockroach extracts. It is hoped that recombinant tropomyosin will be useful for further studies and clinical applications.