曼氏迭宫绦虫裂头蚴可溶性蛋白质的分析

Soluble Proteins of the Plerocercoid of Spirometra mansoni

目的分析曼氏迭宫绦虫裂头蚴可溶性蛋白组分。方法提取曼氏迭宫绦虫裂头蚴总蛋白后,用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和双向凝胶电泳(2-DE)进行分离,转膜后用感染裂头蚴小鼠的血清和健康小鼠血清作为一抗,行蛋白质印迹(Western blotting)分析,比较杂交蛋白点的差异。结果曼氏迭宫绦虫裂头蚴可溶性蛋白经SDS-PAGE分离出33条蛋白带,相对分子质量(Mr)范围在13800～145400,其中Mr26500、Mr37600、Mr88200和Mr130200为高丰度蛋白区带。经Western blotting分析,Mr31600和Mr37600条带为曼氏迭宫绦虫裂头蚴特异性蛋白条带,能被感染小鼠血清识别。裂头蚴总蛋白双向电泳凝胶的蛋白质斑点总数为367个,大部分分布在Mr18840～46800,246个蛋白质斑点的等电点为4.0～7.0,占总数的67%;Western blotting分析显示,30个抗原抗体结合点为特异性抗原斑点。结论曼氏迭宫绦虫裂头蚴可溶性蛋白有2条特异性蛋白带和30个特异性蛋白斑点,以偏酸性为主。

Objective To analyze soluble proteins of the plerocercoid of Spirometra mansoni. Methods The total protein of the plerocercoid of S. mansoni was separated by SDS-PAGE and two-dimensional electrophoresis （2-DE）. Western blotting was performed to find out distinct antigens by sera of plerocercoid-infected mice. Results A total of 33 protein bands were separated by SDS-PAGE （Mr 13 800-145 400）. Four of them were high-abundance proteins with Mr 26 500, Mr 37 600, Mr 88 200, and Mr 130 200, respectively. At least two protein bands of Mr 31 600 and 37 600 reacted with the infected mice sera. 367 protein spots were detected on 2-DE gel, among which about 67% proteins were found within Mr 18 840-46 800 and isoelectric point （pI） 4-7. Western blotting showed 30 antigen spots specifically reacting with sera from mice infected by S. mansoni. Conclusion Two protein bands and thirty protein spots are specific acidic proteins of S. mansoni plerocercoid.