牛皮蝇幼虫蛋白表达谱及一种高表达蛋白的分离纯化

Protein Expression Profiling of Hypoderma Larvae and Purification of A Protein with High Expression Level

研究分析了牦牛皮蝇幼虫蛋白表达谱,为牛皮蝇的基础研究和蛆病防控提供数据。试验提取了从牦牛皮下采集的中华皮蝇(H.sinense)幼虫总蛋白,SDS-PAGE分析幼虫蛋白表达谱,用分子筛和离子交换层析对一种高表达蛋白进行纯化,并对表达谱中差异极大的两个样品进行双向电泳和质谱分析。结果显示,幼虫个体间蛋白表达谱存在不同程度差异,约23%的个体中含有一种分子质量约为78ku的高表达蛋白,纯化并获得了该蛋白质;双向电泳分别获得了312±24和284±13个蛋白点,但对其中的4个蛋白点进行质谱分析,未得到分数高的匹配结果。研究获得了牛皮蝇幼虫蛋白表达谱和纯化的蛋白质,为进一步研究牛皮蝇幼虫的基因表达特征、蛋白质功能等奠定了基础。

The aim of this study was to reveal the protein expression profiling of the Hypoderma larva in-fecting yaks,in order to provide data for the basic research of Hypoderma and the prevention of hypoder-mosis.Total protein of the Hypodermasinense larva collected from subcutaneous tissues of yak was extrac-ted and analyzed by SDS-PAGE.A protein with high was purified using gel exlusion and ion-exchange chromatography and two samples of Hypoderma larvae whose total protein profiling displayed great differ-ence were separated and identified by two-dimensional electrophoresis and mass spectrometry.The results showed that the protein expression profile of larvae varied among individuals,and approximately 23% of the individuals contained a protein with molecular weight of about 78 ku,which was purified by above methods.A total of 312±24 and 284±13 protein spots were obtained respectively by two-dimensional e-lectrophoresis and four of these proteins were analyzed by mass spectrometry,however,none of the pro-teins were identified with high score matches.This study obtained the protein expression profiling of Hy-poderma larvae and purified a protein,laying a foundation for further studies on the gene expression and protein functions of Hypoderma larvae.