| 重组真核质粒pCMV4-rZP3′构建及在小鼠体内的表达 |
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| 引用本文： | 周飞,杨颖,彭景楩.重组真核质粒pCMV4-rZP3′构建及在小鼠体内的表达[J].动物学杂志,2001,36(3):22-27. |
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| 作者姓名： | 周飞 杨颖 彭景楩 |
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| 作者单位： | 中国科学院动物研究所 |
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| 基金项目： | 中国科学院知识创新工程项目，， |
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| 摘 要： | 选取兔透明带ZPC基因序列的部分片段（编码263－415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cDNA，直接插入到克隆质粒PCRR2．1中，构建克隆质粒PCRR2．1－rZP3′，再将靶片段用HindⅢ、X-bal双酶切从克隆质粒PCRR2．1－rZP3′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-rZP3′。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12周以上。
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| 关 键 词： | DNA疫苗 卵透明带 ZPC RT－PCR 重组真核质粒 抗生育 |
| 文章编号： | 0250-3263(2001)03-22-06 |
Construction of the Plasmid pCMV4-rZPC′ DNA Vaccine and Analysis of Its Expression in Mouse |
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| ZHOU Fei,YANG Ying,PENG Jing-Pian.Construction of the Plasmid pCMV4-rZPC′ DNA Vaccine and Analysis of Its Expression in Mouse[J].Chinese Journal of Zoology,2001,36(3):22-27. |
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| Authors: | ZHOU Fei YANG Ying PENG Jing-Pian |
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| 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○ R 2.1 and constructed the PCR○ R 2.1-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. |
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| Keywords: | DNA vaccine Zona pellucida ZPC RT\|PCR |
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