日本血吸虫成虫性别差异蛋白的筛选及鉴定

目的 筛选和鉴定日本血吸虫成虫性别差异蛋白.方法 自日本血吸虫(Schistosomajaponicum,S.japonicum)尾蚴感染的2只家兔颈动脉灌注生理盐水冲虫,分别收集雌雄成虫各约200条.提取雌、雄成虫的水溶性和脂溶性蛋白各300μg,采用双向凝胶电泳技术分别进行分离.银染后扫描成像,用ImageMaster Platinum 2D 5.0软件进行分析,筛选差异蛋白,并予质谱鉴定.结果 雌虫和雄虫的水溶性蛋白分别检出(255±10)、(224±12)个蛋白点,疏水蛋白分别检出(200±11)、(132±8)个蛋白点;鉴定的6个差异蛋白质中,5个雌性表达上调的差异蛋白为硫氧还蛋白,铁蛋白-1重链,泛素结合酶样蛋白Mms2-泛素复合体可溶性结构B链,热休克蛋白10和胞浆脂肪酸结合蛋白突变体H,1个雄性表达上调的差异蛋白为48 kDa组胺受体亚基肽4.结论 获得了日本血吸虫成虫性别差异蛋白质.

Screening and identification of differentially expressed proteins between adult female and male worms of Schistosoma japonicum

Objective To screen and identify differentially expressed proteins between adult female and male worms of Schistosoma japonicum(S.japonicum). Methods Two rabbits infected with the cerearia were peffased with saline in carotid, and approximately two hundred adult female and two hundred male worms of S.japonicums were collected. Approximately 300 μg soluble and hydrophohic proteins of adult female and male worms of S. japonicum were extracted and then the proteins were separated by two-dimensional gel electrophoresis respectively. The analysis using ImageMaster Platinum 2D 5.0 resulted in differentially expressed proteins between adult female and male worms, which were subjected to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and tandem mass spectrometry sequencing of tryptic peptides. Results There were ( 255 ± 10) and ( 224 ± 12 ) spots detected for soluble proteins and (200 ± 11 )and ( 132 ±8 )spots for hydrophobic proteins from adult female and male worms respectively. Six differential proteins were identified, five up-regulated proteins in female worms were thioredoxin,putative ferritin-1 heavy chain,chain B in solution structure of the human ubiquifin-conjugating-enzyme-like protein Mms2-Ubiquitin Complex, heat shock protein 10, cytoplasmic fatty acid binding protein variant H;while only one up-regulated proteins in male worms was identified as 48 kDa histamine receptor subunit peptide 4. Conclusion Several differentially expressed proteins between female and male worms of S.japonicum were recognized through screening and identifying differential proteins between female and male worms of S. japonicum.