【摘要】 利用RT-PCR技术从黑龙江省猪源旋毛虫获得了Ts43基因,并克隆入pcDNA3.1-CT-GFP真核表达载体中构建重组质粒,用该重组质粒在脂质体介导下转染Vero E6细胞,GFP标签证明质粒DNA成功转染到细胞中并得以表达,通过Western-blot分析,细胞裂解液样品中有1条约66 ku的条带,可被小鼠旋毛虫阳性血清所识别,与预计大小一致,说明,真核表达载体pcDNA3.1-CT-GFP中的Ts43基因在VeroE6细胞中获得了表达,表达产物具有抗原性。更多还原

【Abstract】 The Ts43 gene of Trichinella spiralis originated from pigs in Heilongjiang Province was amplified by RT-PCR and subcloned into pcDNA3.1-CT-GFP vector to construct recombinant plasmid pcDNA3.1-CT-GFP-Ts43.Sequence analysis revealed that the Ts43 gene was correctly inserted into the pcDNA3.1-CT-GFP.Then,the pcDNA3.1-CT-GFP-Ts43 plasmid was transfected into Vero E6 cells mediated by Lipofectamine 2000.GFP showed that the recombinant plasmid was expressed in Vero E6 cells.Western-blot analysis showed that a band approximately 66 ku in size in Vero E6 cells lysate was recognized by the mouse serum against T.spiralis,indicating that the Ts43 protein expressed transiently in the cells had antigenicity.更多还原