菲律宾蛤仔过敏原原肌球蛋白的鉴定与分子克隆

目的了解菲律宾蛤仔中过敏原的情况,对其主要过敏原进行鉴定和分子克隆。方法采用聚丙烯酰胺凝胶电泳和免疫印迹验证原肌球蛋白,双向电泳对蛋白等电点进一步确定。利用差示扫描量热法对蛋白的热性能测定以及蛋白克隆和测序来分析原肌球蛋白。结果菲律宾蛤仔原肌球蛋白的分子量在37 k Da左右,等电点为5.1,热稳定性较强。原肌球蛋白的基因序列全长为855 bp,编码284个氨基酸,对序列进行同源对比,相似性较高。结论本实验证实了原肌球蛋白为菲律宾蛤仔的过敏原,为认识菲律宾蛤仔过敏原提供基础数据。

Identification and molecular cloning of the allergen tropomyosin from Ruditapes philippinarum

Objective To investigate the Ruditapes philippinarum allergen, and identify and clone the main allergens. Methods Tropomyosin was identified by polyacrylamide gel electrophoresis and immunoblotting, and isoelectric point was further analyzed by two dimensional electrophoresis. Thermal stability analysis was determined by differential scanning calorimetry, and tropomyosin was analyzed by cloning and sequencing of proteins. Results The major allergen protein tropomyosin with the molecular weight 37 kDa was extracted. The isoelectric point was 5.1. The tropomyosin of R.philippinarum was stable in the process of thermal treatment. The full-length cDNAs encoding tropomyosin was composed of 855 bp coding for 284 amino acid residues. The similarity of sequence was high through homologous comparison. Conclusion The tropomyosin is the allergen of R.philippinarum. This research is helpful to provide the basic date to understand R.philippinarum allergen.