| 番鸭呼肠孤病毒σC基因重组腺病毒载体的构建和鉴定 |
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| 引用本文： | 林锋强,程晓霞,王劭,朱小丽,王锦祥,陈仕龙,陈少莺.番鸭呼肠孤病毒σC基因重组腺病毒载体的构建和鉴定[J].福建农业学报,2016(10):1015-1019. |
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| 作者姓名： | 林锋强 程晓霞 王劭 朱小丽 王锦祥 陈仕龙 陈少莺 |
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| 作者单位： | 福建省农业科学院畜牧兽医研究所/福建省畜禽疫病防治工程技术研究中心,福建 福州,350013 |
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| 基金项目： | 国家自然科学基金项目(31172334)，福建省自然科学基金项目(2016J01137) |
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| 摘 要： | 应用RT-PCR方法扩增出番鸭呼肠孤病毒(Muscovy duck reovirus,MDRV)的σC基因,通过XhoⅠ+HindⅢ双酶切后把该基因插入到经过同样双酶切的穿梭质粒pShuttle-CMV载体中。重组穿梭载体经过双酶切和PCR鉴定后进行测序,序列测定正确,同源性为99.8%。将获得的重组穿梭质粒与腺病毒骨架质粒共转染293细胞后获得重组腺病毒。PCR和RT-PCR鉴定的结果证明σC蛋白基因已成功插入到重组腺病毒载体中。
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| 关 键 词： | 番鸭呼肠孤病毒 σC基因 重组腺病毒载体 |
Construction and Certification of Recombinant Adenovirus Vector forσC Gene in Muscovy Duck Reovirus |
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| Abstract: | TheσC gene of muscovy duck reovirus (MDRV)was amplified by RT-PCR and cloned into pShuttle-CMV plasmid after the amplicon was digested by XhoI+Hind III enzyme.Sequencing analysis confirmed the gene to beσC.Subsequently,the recombinant plasmid pShuttle-CMV-σC and the adenovirus backbone vector,pAdEasy-1 , were co-transfected into 293 cells to construct a recombinant adenovirus.The obtained recombinant adenovirus pAd-σC was positively identified by PCR and RT-PCR.The information obtained could be used for genetic engineering a vaccine against MDRV. |
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| Keywords: | muscovy duck reovirus σC gene recombinant adenovirus vector |
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