摘要
为研究旋毛虫Ts87基因的功能,通过RNA干扰途径抑制目的基因的表达。针对Ts87基因序列设计并合成特异性dsRNA、siRNA,通过浸泡方式分别将dsRNA、siRNA导入肌幼虫体内,利用实时荧光定量PCR分析Ts87基因及旋毛虫管家基因GAPDH的表达。此外,测定RNA干扰作用下的肌幼虫体外存活率的变化。根据实时荧光定量PCR结果,dsRNA及siRNA均能有效抑制Ts87基因mRNA的表达。此外,两种RNA能够在一定程度上降低肌幼虫体外存活率。实验表明,RNA干扰技术能够有效抑制旋毛虫功能基因Ts87表达,从而为研究旋毛虫基因的功能提供一种新的有效途径。
To investigate the functions of Ts87 gene of Trichinella spiralis, we blocked its expression with interference of RNA. Based on the sequence of Ts87 gene, specific dsRNA and siRNA were designed, synthesized and delivered into T. spiralis larvae by soaking. The expression levels of Ts87 and house-keeping gene GAPDH of T. spiralis mRNA were quantified using real time PCR. Besides, the survival rates of larvaes treated by RNAi in vitro were detected. The results showed that the expression of Ts87 mRNA in larvaes exposed to dsRNA or siRNA was reduced significantly, and the survival rates of larvaes in vitro were also observed reduced with the interference of two kinds of RNAs in some extent. Expression of functional gene Ts87 of T. spiralis can be effectively interfered by siRNA, and it paved a new strategies to investigate the gene function of T. spiralis.
出处
《寄生虫与医学昆虫学报》
CAS
2008年第1期26-33,共8页
Acta Parasitologica et Medica Entomologica Sinica
基金
国家自然科学基金项目(No.30571626)
北京市自然科学基金项目(No.5062005)
北京市市属市管高等学校"拔尖创新人才"资助项目
国家教育部博士点基金项目(No.20060025003)
北京市教委科技发展重点项目(KZ200810025009)
