| CAG和MLP启动子控制的重组禽腺病毒构建 |
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| 引用本文： | 张明明,秦爱建,庄骁颖,王小辉.CAG和MLP启动子控制的重组禽腺病毒构建[J].畜牧与兽医,2010,42(3). |
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| 作者姓名： | 张明明 秦爱建 庄骁颖 王小辉 |
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| 作者单位： | 扬州大学兽医学院,江苏,扬州,225009 |
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| 摘 要： | 根据鸡胚致死孤儿病毒(CELOV)的序列设计引物,通过PCR方法扩增了禽腺病毒FAVI-JS株的晚期启动子(MLP),序列与CELOV的MLP同源性为99%。分别构建了MLP和商品化启动子CAG控制的表达增强型绿色荧光蛋白(eGFP)基因的禽腺病毒转移载体。通过载体与禽腺病毒FAVI-JS株在原代鸡胚肾细胞内进行同源重组,分别获得了MLP和CAG控制的表达eGFP的重组禽腺病毒,为探讨不同启动子对禽腺病毒活载体疫苗免疫效果的影响提供了条件。
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| 关 键 词： | 巨细胞病毒增强子和鸡β肌动蛋白启动子 晚期启动子 重组 禽腺病毒 |
Construction of recombinant avian adenovirus controlled by CAG and MLP promoters |
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| Abstract: | The major late promoter(MLP) of avian adenovirus FAVI-JS was amplified by PCR with the pair of primers designed based on the nucleotide sequence of chicken embryo lethal orphan virus(CELOV).And the sequence of MLP had high homology(99%) with that of CELOV.The two plasmids pFAMLP-eGFP and pFACAG-eGFP were constructed as transfer vectors with MLP-eGFP-BGH PA or CAG-eGFP-BGH PA inserted between the right arm and the left arm for recombination.The recombinant avian adenoviruses rFAMLP-eGFP and rFACAG-eGFP were successfully constructed by the method of recombination between pFAMLP-eGFP or pFACAG-eGFP and FAVI-JS.The study provided a basis for investigating the influence of different promoters on the immune efficiency of recombinant avian adenovirus. |
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| Keywords: | CMV early enhancer/chicken β actin(CAG) promoter major late promoter(MLP) recombination avian adenovirus |
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