广州管圆线虫抗原IF的中间纤维蛋白组织来源分析和定位

目的 对广州管圆线虫抗原IF进行中间纤维蛋白家族分类和细胞组织定位.方法 IFTG诱导重组基因pET-IF在大肠杆菌中表达广州管圆线虫抗原IF,用组氨酸亲和层析纯化表达产物,用Western blot方法鉴定抗原IF的中间纤维蛋白类型;小鼠皮下多点注射法制备抗IF抗体血清,用免疫组织化学方法检测抗原IF在虫体组织和细胞上的定位.结果 广州管圆线虫抗原IF在体外成功表达并纯化,其属于中间纤维蛋白家族的角蛋白,定位于广州管圆线虫成虫肠管壁,存在于细胞质中.结论 广州管圆线虫抗原IF分布于虫体肠管壁,同时从组织学角度证明广州管圆线虫存在中间纤维结构.

Identification and tissue localization of intermediate filament protein in Angiostrongylus Cantonensis

Objective To identify the type of the intermediate filament(IF)protein of Angiostrongylus cantonensis and analyze its tissue localization.Methods Recombinant pET-IF of antigen IF was expressed in E.coli with IPTG induction,and the expression products were purified by His.Bind column and identified for determining the type of the IF protein by Western blotting.Anti-IF antibody was prepared by multi-spot subcutaneous injection into mouse and used to detect the tissue slices of A.cantonensis by immunohistochemical analysis.Results The antigen IF were correctly expressed and purified,and identified as a keratin located in the intestine wall and cytoplusma.Conclusion The antigen IF is distributed in the intestine wall of A.cantonensis.