大劣按蚊感染约氏疟原虫血淋巴相关蛋白的筛选和鉴定

Screening and identification of the related hemolymph protein of Anopheles dirus against Plasmodium yoelii infection

目的筛选并鉴定大劣按蚊感染约氏疟原虫后血淋巴的相关蛋白,为进一步的功能研究和蚊先天性免疫研究奠定基础。方法以大劣按蚊-约氏疟原虫为实验模型,利用二维电泳技术分离和比较饱吸约氏疟原虫感染的鼠血和正常鼠血后24 h的大劣按蚊血淋巴蛋白,获得差异蛋白,并对差异蛋白进行N端氨基酸测序和生物信息学分析,从而推测差异蛋白的功能。结果获得清晰二维电泳图,并筛选获得37个差异蛋白点。对其中比较明显的差异蛋白1进行氨基酸测序后发现,该蛋白可能为一翻译延伸因子。结论成功分离和筛选获得了大劣按蚊感染约氏疟原虫血淋巴相关蛋白,并进行了初步鉴定。

Objective To screen and identify the related protein of Anopheles dirus against Plasrnodiurn yoelii infection. Methods Proteins from hemolympb of An, dirus infected and uninfected with P. yoelii were separated with twodimensional pol3,acrylamide gel electrophoresls （2-DE） respectively. Then the gel were stained by Coomassie brilliant blue and analysed by PDQuest 2D Elite analysis software. The differentially expressed proteins were obtained and the N termi hal amino acid sequencing were carried out. At last the function 6f the differentially expressed protein were forecasted by NCBInr database searching. Results The 2 DE patterns with high resolving power and reproducibility were obtained, 37 differentially expressed protein spots were found by comparing the two gels. Base on the result of amino acid sequencing, differentially expressed protein 1 is likely a kind of translation elongation factor. Conclusion The related protein of An. dirus against P. yoelii infection were successfully separated and identified primarily.