中文摘要：

为构建犬新孢子虫NcAMA1基因重组腺病毒穿梭质粒，并在真核细胞中表达，以重组质粒pVAX1-NcAMA1为模板，PCR扩增了NcAMA1基因，以此构建了pMD18T—NcAMA1重组克隆质粒；对该重组克隆质粒进行双酶切鉴定后，将其亚克隆至腺病毒穿梭载体pCR259中。应用脂质体介导转染法将经PCR鉴定和酶切鉴定正确的pCR259-NcAMA1重组穿梭质粒转染293细胞，应用IFAT和Western—blot技术检测了AMA1基因在293细胞中的表达情况。结果显示，扩增的NcAMA1基因长度为1695bp，构建的重组腺病毒穿梭质粒pCR259-NcAMA1能在293细胞中获得瞬时表达，表达蛋白的分子质量约为68ku。

英文摘要：

To construct recombinant adenovirus shuttle plasmid with Neospora caninum NcAMA1 gene and to express the recombinant plasmid in eukaryotic cells, using the constructed eukaryotic recombi- nant plasmid pVAX1-NcAMA1 as a template, NcAMA1 gene was amplified by PCR, and the recombinant plasmid pMD18T-NcAMA1 was consturcted. The digested NcAMA1 gene was subcloned into adenovirus shuttle vector pCR259 digested with the same enzymes. The NcAMA1 recombinant adenovirus shuttle plasmid pCR259-NcAMA1 was then transfected into 293 cells by liposome. The expressed product in 293 cells was analyzed by IFAT and Western-blot. In result,the amplified gene NcAMA1 was 1 695 bp in size, the recombinant adenovirus shuttle plasmid pCR259-NcAMA1 was constructed successfully, and the recombinant plasmid transiently expressed in 293 cells,and the recombinant protein was about 68 ku in a mo- lecular mass.