日本囊对虾原肌球蛋白的原核表达、抗体制备以及组织表达分析

Prokaryotic expression, antibody preparation and tissue expression identification of tropomyosin in Marsupenaeus japonicas

食物过敏是人类常见的一种过敏性疾病,主要由食物中的蛋白质引起。原肌球蛋白(tropomyosin,TPM)是食用虾蟹等甲壳动物引起过敏反应的主要致敏物质。研究从日本囊对虾肌肉组织中克隆了TPM基因,进行了原核表达和蛋白质纯化,并进一步制备了相应的抗体。Western-blotting分析表明原肌球蛋白在日本囊对虾组织中普遍表达,并且肌肉组织中表达量最高而鳃和皮肤组织中表达量最低。研究结果为对虾过敏性疾病的诊断和治疗以及脱敏食品开发等奠定了理论基础。

Food allergy is a representative human allergic disease, which is mainly caused by the protein in food. Up to date, however, there is only a few reports on aquatic food allergy. Tropomyosin（TPM） is the major allergen of eating seafood like shimp and crab to cause hypersensitivity. In this investigation, a TPM gene was cloned from the muscle tissue of shrimp, Marsupenaeus japonicus. Then the TPM was expressed in Escherichia coli and purified. Subsequently the specific antibody was raised using the purified fusion protein （GST-TPM）. As revealed by Western-blotting, the TPM was ubiquitously expressed in shrimp tis-sues and with the highest expression in the muscle and the lowest in gill and skin. These results have laid the foundation for the specific diagnosis and immunotherapy in shrimp hypersensititivity disease as well as the study on food desensitization.