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目的分离、鉴定中国大陆日本血吸虫不同地理株间成虫差异表达蛋白。方法分别收集、制备日本血吸虫不同地理株成虫可溶性蛋白,经固相pH梯度双向凝胶电泳分离,凝胶银染,并用PDQuest8.0凝胶图像分析软件进行比较分析,筛选出差异表达蛋白点并采用基质辅助激光解析离子飞行时间质谱仪进行鉴定。结果湖南株、江西株和江苏株雄虫分别检出698±10、650±19、629±23个蛋白点,雌虫分别检出670±12、682±22、625±28个蛋白点,约90%蛋白点分子量处于20～90kD范围内,蛋白等电点在5～8之间。不同地理株雌虫与雄虫分别两两比较,雄虫发现14个差异点,雌虫发现18个差异点。对其中7个雄虫差异表达蛋白及3个雌虫差异表达蛋白经MALDI-TOF-MS鉴定分析,获得其肽质量指纹图谱、等电点、分子量等相关信息。7个雄虫差异表达蛋白分别为:SJCHGC00821蛋白、SJCHGC00475蛋白、3-磷酸甘油醛脱氢酶、谷胱甘肽转移酶片段、谷胱甘肽-S-转移酶、D-核酮糖-5-磷酸-3-表异构酶、G蛋白α亚基AgGq1,3个雌虫表达蛋白分别为:预测蛋白、GAF型传感器信号转导组氨酸激酶、组织蛋白酶B样半胱氨酸蛋白酶前体。结论中国大陆日本血吸虫不同地理株虫体间蛋白质存在差异,部分蛋白表达差异与虫体对吡喹酮敏感性密切相关。
Objective To isolate and identify differentially expressed proteins of adult worms from different geographic strains of Schistosoma japonicum in mainland China. Methods Soluble proteins of adult schistosomiasis strains were collected and separated by two-dimensional gel electrophoresis (SDS-PAGE) and gel-silver staining. The proteins were analyzed by PDQuest 8.0 gel image analysis software, and the differentially expressed protein spots were screened Matrix-assisted laser desorption ionization time-of-flight mass spectrometry was used for identification. Results The Hunan, Jiangxi and Jiangsu strains detected 698 ± 10,650 ± 19,629 ± 23 protein spots respectively, and the females detected 670 ± 12,682 ± 22,625 ± 28 protein spots respectively The molecular weight of 90% protein spots is in the range of 20-90kD, and the isoelectric point of proteins is between 5-8. Different geographical strains of male and female were compared with each other, male 14 found differences, 18 differences found in the female. The differentially expressed proteins of seven male and three female differentially expressed proteins were identified by MALDI-TOF-MS, and the related information of peptide mass fingerprinting, isoelectric point and molecular weight were obtained. The differentially expressed proteins of the 7 male species were SJCHGC00821 protein, SJCHGC00475 protein, glyceraldehyde-3-phosphate dehydrogenase, glutathione transferase fragment, glutathione-S-transferase, D- - phospho-3-epimerase, G protein α subunit AgGq1, 3 female expressed proteins were: predicted protein, GAF sensor signal transduced histidine kinase, cathepsin B-like cysteine protease before body. Conclusion There are differences in the proteins among different geographical strains of Schistosoma japonicum in mainland China. The differences of some protein expressions are closely related to the sensitivity of parasites to praziquantel.