感染约氏疟原虫前后大劣按蚊血淋巴的差异表达蛋白分析

目的获得并分析感染约氏疟原虫前后大劣按蚊血淋巴的差异表达蛋白.方法利用二维电泳方法分别分离感染约氏疟原虫前后的大劣按蚊血淋巴蛋白,进行考马斯亮蓝染色,并利用PDQuest 2D Elite软件分析和比较凝胶结果;对感染约氏疟原虫前后的差异表达蛋白进行质谱分析,获得肽质量指纹图谱;利用Mascot软件系统,在Swiss-Prot等数据库中检索和分析所获得的肽质量指纹图谱.预测差异表达蛋白的生物学特性和在卵囊黑化中的功能.结果获得了分辨率和重复性均较好的二维电泳图,感染组凝胶可见120个蛋白点,正常组凝胶可见95个蛋白点,二者比较分析后发现有37个差异蛋白点;质谱分析获得3个肽质量指纹图谱,于蛋白数据库检索后未发现匹配分值很高的相关蛋白,其中差异蛋白点1与冈比亚按蚊一蛋白具有较高的同源性.结论获得了感染约氏疟原虫前后大劣按蚊血淋巴的差异蛋白和部分差异蛋白的相关信息,这为从蛋白水平认识蚊媒和疟原虫的相互关系和更深入地利用大劣按蚊-约氏疟原虫模型开展卵囊黑化的相关研究奠定了基础.

Analysis of differentially expressed proteins from hemolymph of Anopheles dirus in-fected and uninfected with Plasmodium yoelii

Objective To investigate the differentially expressed proteins from hemolymph of Anopheles dirus infected and uninfected with Plasmodium yoelii. Methods Proteins from hemolymph of Anopheles dirus infected and uninfected with Plasmodium yoelii were separated with two-dimensional polyacrylamide gel electrophoresis (2-DE) respectively. Then the gel were stained by Coomassie brilliant blue and analyzed by PDQuest 2D Elite analysis software. Mass spectrometry and NCBInr database searching were carried out to analyze and forecast the biologic character and function in oocyst melanization of the differentially expressed proteins. Results The 2-DE patterns with higher resolving power and reproducibility were obtained. There were 120 protein spots in the gel of infected group and 95 protein spots in the gel of uninfected group. A total of 37 differentially expressed protein!spots were found by comparing the two gels. We got 3 peptide mass fingerprints (PMF) by mass spectrometry. There was no protein matching highly with the 3 analyzed proteins in the NCBInr database. However, an unknown protein of Anopheles gambiae homologous with the differentially expressed protein spot 1 was found in the database. Conclusion We got the differentially expressed proteins from hemolymph of Anopheles dirus infected and uninfected with Plasmodium yoelii. Some of their information were also obtained. Our study will help to understand the relationship between Anopheles and Plasmodium on protein level and carry out the further study on oocyst melanization using the model of Anopheles dirus-Plasmodium yoelii.