期刊论文

Liu, Y.

中文

中国寄生虫学与寄生虫病杂志

1000-7423

2011

29

1

16-20

国家自然科学基金（No.30570952） 湖南省卫生厅项目（2009） 湖南省科技项目（2010）

本校为其他机构

目的观察日本血吸虫表膜特异结合肽合成基因(ZLW)构建的ZLW/pEGFP-C2重组质粒体外转染日本血吸虫童虫的效率,了解其抗虫作用.方法用0.75%二甲基亚砜(DMSO)和高浓度质粒浸泡法,使用重组质粒ZLW/pEGFP-C2转染机械转化的日本血吸虫脱尾童虫,以瞬时表达的增强型绿色荧光蛋白(EGFP)基因为报告基因,在倒置荧光镜下观察虫体.抽提转染后培养48h虫体的总RNA和全虫蛋白,分别用RT-PCR和蛋白质印迹(Western blotting)检测ZLW基因和EGFP基因在虫体内的表达情况.于转染后培养24,48,72和96h,用美蓝染色法鉴别虫体存活情况,并计数.以上试验均设空质粒组和Tris-HCl缓冲盐溶液(TBS)对照组.结果镜下观察结果显示,转...

OBJECTIVE: To study the efficiency of ZLW/pEGFP-C2 plasmid transfection into Schistosoma japonicum schistosomula and observe its in vitro effect of anti-schistosomula. METHODS: Recombinant plasmid ZLW/pEGFP-C2 was transfected into mechanically transformed schistosomula by immersion in 0.75% DMSO and high concentration of plasmid. Enhanced green fluorescent protein (EGFP) transfected cells were observed under inverted fluorescence microscope. At 48 hours after culture, total RNA and proteins from transfected schistosomula were extracted, and the presence of the transgenes (ZLW and EGFP) in schi...