中国明对虾血浆蛋白质组双向电泳技术的建立与主要高丰度蛋白点的质谱鉴定

中国明对虾的血浆脱盐处理后,通过固相pH梯度胶条等电聚焦、SDS-PAGE垂直电泳对蛋白质进行分离,对不同pH范围IPG胶条、脱盐方式、上样量等进行了优化,并分别利用硝酸银和胶体考染方法进行染色,应用PDQuest软件对图谱进行了初步分析,采用基质辅助激光解析电离-飞行时间质谱法对主要的高丰度蛋白点进行了质谱鉴定。结果显示,中国明对虾血浆在pH 4~7范围、18 cm的2-DE胶上得到了很好的分离,上样量为150 μg左右,采用银染可以获得较好的分析胶图谱；而上样1 000 μg左右,采用胶体考染方法能获得较好的制备胶图谱；通过MALDI-TOF/TOF MS鉴定的11个最主要的高丰度蛋白点都是血蓝蛋白。建立并优化了中国明对虾血浆蛋白质组学的双向电泳技术体系,为进一步开展对虾等甲壳动物的血浆蛋白质组学研究奠定了基础。

Establishment of two-dimensional gel electrophoresis technique for Fenneropenaeus chinensis plasma and identification of major high-abundance proteins by MALDI-TOF/TOF MS

Chinese shrimp Fenneropenaeus chinensis is one of the most economically valuable and widely cultured species in China.The hemolymph is the source of its immunity to disease and environmental factors,which invariably involves protein components.To better understand the physiology of F.chinensis and the major protein components expressed during its life cycle,two proteomics approaches,two-dimensional gel electrophoresis(2-DE) in conjunction with MALDI-TOF/TOF MS,were employed to analyze its hemolymph proteins...