大劣按蚊与烟草天蛾丝氨酸蛋白酶的交叉免疫活性

Cross-immunoreactivity between serine proteases from Anopheles dirus and Manduca

目的探讨大劣按蚊与烟草天蛾丝氨酸蛋白酶之间是否存在交叉免疫反应性。方法首先利用生物信息学方法分析不同昆虫间丝氨酸蛋白酶的氨基酸序列的保守性,尤其是大劣按蚊与烟草天蛾间丝氨酸蛋白酶的序列相似性;然后,PinPoint Xa-1表达载体对大劣按蚊丝氨酸蛋白酶进行体外原核表达,利用Western blot方法检测表达产物与烟草天蛾丝氨酸蛋白酶抗血清之间的免疫反应性。结果生物信息学分析结果显示,不同昆虫间(包括大劣按蚊与烟草天蛾)丝氨酸蛋白酶的氨基酸序列具有较高保守性;成功进行了大劣按蚊丝氨酸蛋白酶的体外原核表达,western blot结果显示,烟草天蛾丝氨酸蛋白酶抗血清能够能够识别表达产物。结论大劣按蚊与烟草天蛾丝氨酸蛋白酶具有交叉免疫活性,利用烟草天蛾丝氨酸蛋白酶抗血清研究大劣按蚊丝氨酸蛋白酶是可行的。这将为下一步从蛋白水平研究大劣按蚊丝氨酸蛋白酶的生物学特性奠定基础。

Objective To investigate the cross-immunoreactivity between the serine proteases derived from Anopheles dirus and Manduca. Methods Bioinformatic analysis were carried out to determine the conservation of serine protease amino acid sequences among different insects, especially its similarity of the 2 above-mentioned insects. Serine protease of Anopheles dirus was expressed in E. coli by PinPointTM Xa-1 T-vector. Western blotting was used to examine the cross-immunoreactiviy between the expression product and the antiserum against serine protease of Manduca. Results High similarity was found in serine protease amino acid sequences of different insect, including Anopheles dirus and Manduca. Serine protease of Anopheles was successfully expressed in E. coli. Western blot analysis showed that the expression product was recognized by the antiserum. Conclusion There is cross-immunoreactivity between serine protease of Anopheles dirus and Manduca. So it is feasible to study serine protease of Anopheles dirus with antiserum against Manduca serine protease.