【摘要】 选取兔透明带ZPC基因序列的部分片段 (编码 2 63 4 1 5位的氨基酸 ,rZP3′)作为靶抗原 ,构建兔ZP3′基因疫苗 ,研究其阻断受精的效果。提取成熟雌兔卵巢的总RNA ,RT PCR克隆兔透明带ZPC的cD NA ,直接插入到克隆质粒PCR○R 2 1中 ,构建克隆质粒PCR○R 2 1 rZPC′,再将靶片段用HindⅢ、X bal双酶切从克隆质粒PCR○R 2 1 rZPC′切下 ,1 %的低熔点琼脂糖电泳回收 ,亚克隆到真核表达质粒pCMV4中 ,构建真核表达质粒pCMV4 rZPC′。将质粒直接注射到活体小鼠的大腿内侧肌肉中 ,在mRNA水平上观察到质粒能在肌肉细胞中正确表达 ,并且其表达可以维持 1 2周以上。更多还原

【Abstract】 Selecting the partial sequence of rabbit zona pellucida protein C (Amino Acid No.263\|415, rZPC′) as the target, the pCMV4\|rZPC′ gene vaccine was constructed. The total\|RNA was extracted from the ovary of sexually matured healthy female rabbit (New Zealand) and the rZPC′ cDNA was amplified by RT\|PCR. The product of RT\|PCR was directly inserted into the cloning vector PCR○R2.1 and constructed the PCR○R2.1\|rZPC′. Then the vector was digested by Hind Ⅲ, X\|bal and the inserts were subcloned into plasmid pCMV4 to construct pCMV4\|rZPC′ DNA vaccine. Analysis by RT\|PCR demonstrated that intramuscular injection of mouse with pCMV4\|rZPC′ vector resulted in an expression in mice muscle and the expression could be up to 12 weeks.更多还原