美洲大蠊新过敏原的克隆，表达及致敏性研究

American cockroach （Periplaneta americana）is an important insect for drug development. However, cockroach is also a major indoor allergen. At present, many allergens have been identified from American cockroach. Our previous study found another six novel potential allergens from American cockroach by allergomics method. The current project firstly constructs American cockroach cDNA library and clones these novel allergens by the primes designed according to the partial amino acid sequences of allergens identified by MALDI-TOF/TOF MS. Then, these novel allergens are expressed in both E.coli and eukaryotic insect cell systems. The allergenicities of these allergens are tested by ELISA, immune-blotting, basophils activation test，skin prick and T cell proliferation assay. The identified novel American cockroach allergens can give important allergens information for drug development of American cockroach. Moreover, these allergens can be developed to be diagnosis reagent and vaccine for cockroach allergy.

蟑螂，特别是美洲大蠊为我国重要药用动物资源。然而蟑螂本身是仅次于尘螨的第二大室内过敏源。目前已经从美洲大蠊中鉴定出多个过敏原成分。对美洲大蠊过敏蛋白组学前期研究在美洲大蠊中发现出6个新的潜在过敏原蛋白。本项目首先构建美洲大蠊cDNA文库，根据质谱鉴定得到的新过敏原部分氨基酸序列设计引物，调取美洲大蠊新过敏原基因。在大肠杆菌和杆状病毒-昆虫表达系统中表达并纯化，得到新过敏原蛋白。通过ELISA，免疫印迹，嗜碱性粒细胞激发试验，皮肤点刺试验研究新过敏原的致敏性，明确美洲大蠊新过敏原。从而对美洲大蠊过敏原成分信息有个全面的认识，为今后美洲大蠊药物的开发和利用奠定基础。同时鉴定出的过敏原蛋白本身可以开发成诊断试剂和脱敏疫苗用于蟑螂过敏的诊断和治疗。

蟑螂，特别是美洲大蠊为我国重要药用动物资源。然而蟑螂本身是仅次于尘螨的第二大室内过敏源。目前已经从美洲大蠊中鉴定出多个过敏原成分。本研究通过过敏蛋白组学方法在美洲大蠊中发现出6个新的潜在过敏原蛋白。其中对于Per a5进行了基因克隆，得到全长cDNA，并在大肠杆菌和杆状病毒-昆虫表达系统中表达并纯化，得到新过敏原蛋白。通过ELISA，免疫印迹，嗜碱性粒细胞激发试验研究新过敏原Per a5的致敏性。本项目共发表第一标注SCI收录论文5篇。