日本血吸虫单性感染雄虫和双性感染雄虫差异表达蛋白的筛选与鉴定  

Screening and Identification of Differential Expression Proteins Between Male Worms of Single-sex and Bisexual Infection in Schistosoma japonicum by Two-dimensional Electrophoresis

作　　者：戴橄[1] 汪世平[1] 余俊龙[1] 徐绍锐[1] 彭先楚[1] 刘雪琴[1] 周松华[1] 刘芬[1] 

机构地区：[1]中南大学湘雅医学院血吸虫病研究室,长沙410078

出　　处：《生物化学与生物物理进展》2007年第3期283-291,共9页Progress In Biochemistry and Biophysics

基　　金：国家"十.五"重大科技专项课题(2002AA2Z3343);国家高技术研究发展计划(863)(2004AA2Z3530);湖南省"十一五"重大专项(2006SK1001);湖南省重点学科建设专项经费联合资助~~

摘　　要：采用双向凝胶电泳和质谱技术对日本血吸虫单性感染雄虫和双性感染雄虫蛋白质表达谱进行分析,并在mRNA水平进行验证;观察差异表达SJCHGC蛋白重组真核表达质粒pEGFP-C1/SJCHGC在COS-7细胞中的表达和亚细胞定位,并分析其表达产物的抗原性.成功鉴定了9个日本血吸虫雌雄合抱差异表达蛋白,其中在日本血吸虫雄虫合抱前表达上调的蛋白质有6个;而有3个蛋白质在日本血吸虫雄虫合抱后表达明显上调.大多数差异蛋白功能涉及血吸虫的生长发育、生殖、营养、运动、信号传递等过程.重组质粒pEGFP-C1/SJCHGC融合基因真核表达载体在真核细胞COS-7中获得了表达,可用荧光显微镜直接观察其表达情况及亚细胞定位,细胞所表达的融合蛋白具有血吸虫抗原性.研究结果为揭示日本血吸虫雌雄合抱机制、研制抗血吸虫雌雄合抱疫苗提供了理论依据.Two-dimensional （2D） gel electrophoresis followed by mass spectrometry was utilized to compare the protein expression between male worms of single-sex infection and male worms of bisexual infection in Schistosoma japonicurn. To observe the in vitro expression and subcellular localization of recombinant eukaryotic expression plasmid pEGFP-C 1/SJCHGC in COS-7 cells and analyze the antigenicity of the products expressed. 9 pairing-related proteins were identified and further validated by semi-quantitative RT-PCR analysis. Most of the candidate proteins have been evident to be somehow associated with various aspects ofS. japonicum such as cell growth, development, motility, reproduction, nutrition and signal transmission. The SJCHGC gene was expressed in the transfected COS-7 cells. The expression and subcellular localization of the fusion protein were analyzed by fluorescence microscopy. The proteins expressed show the antigenicity of Schistosoma japonicum. These results provide the basis for elucidating the Schistosome mechanism of development and reproduction, and may permit the identification of protein candidates for the development of novel vaccine or new targets for drug development against schistosomiasis.

关 键 词：日本血吸虫 雌雄合抱 蛋白质组学 双向电泳(2-DE) 基因克隆 绿色荧光蛋白 COS-7细胞 

分 类 号：R383[医药卫生—医学寄生虫学]