中文摘要：

PCR扩增牛源犬新孢子虫NcAMAl基因。克隆至pMD18-T simple载体；将鉴定正确的NcAMA1基因亚克隆至pCR259腺病毒穿梭载体，构建重组腺病毒穿梭质粒pCR259-NcAMA1；依次转化HighQ-1 Transpose—Ad 294和HighQ-1感受态细胞，构建NcAMA1基因重组腺病毒表达质粒T294-NcAMA1；Pac I酶线性化后，脂质体介导转染至QBI—HEK293细胞，包装Ad5-NcAMA1重组腺病毒。结果表明，获得Ad5-NcAMA1重组腺病毒滴度为5．6×10^9TCID50／mL；经PCR检测到重组腺病毒NcAMA1基因；经IFAT和Western blot检测NcAMA1基因在QBI—HEK293细胞中获得表达．表达蛋白相对分子质量为68000，具有较好的反应原性。本试验成功构建了Ad5-NcAMA1重组腺病毒，为牛源犬新孢子虫NcAMA1基因重组腺病毒载体疫苗的研制奠定基础。

英文摘要：

NcAMA1 gene of bovine Neospora caninum was amplified by PCR,pCR259-NcAMA1 was constructed,then transformed into HighQ 1 Transpose-AdTM 294 and HighQ-1TM competent cell,T294-NcAMA1 were constructed. Coated with liposome, T294 NcAMA1 was transfected into QBI-HEK293 cells to package recombinant adenovirus Ad5-NcAMA1. Recombinant adenovirus NcAMA1 gene was detected by PCR. The expression of NcAMA1 gene in QBI-HEK293 cells was detected by IFAT and Western blot. Result showed that the recombinant adenovirus Ad5-NcA- MA1 was successfully packaged into 293 cells. The protein expressed by AdS-NcAMA1 was 68 000 with had good reactogenicity. The titer of recombinant adenovirus AdS NcAMA1 was 5. 6 ×109 TCID50/mL. It laid a solid foundation for the development of recombinant adenovirus vaccine against Neospora caninum with NcAMA1 gene.