| NIPSNAP3A短发夹RNA腺病毒载体的构建及病毒包装 |
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| 引用本文： | 俞华吉,闫永红,唐刘君,杨晓明,王晓辉,汪思应.NIPSNAP3A短发夹RNA腺病毒载体的构建及病毒包装[J].生物技术通讯,2012,23(4):485-487,492. |
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| 作者姓名： | 俞华吉 闫永红 唐刘君 杨晓明 王晓辉 汪思应 |
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| 作者单位： | 1. 安徽医科大学病理生理学教研室,安徽合肥,230032
2. 军事医学科学院放射与辐射医学研究所,北京蛋白质组研究中心,北京 102206 |
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| 基金项目： | 国家自然科学青年基金(30900546);国家重点基础研究发展计划(2011CB910600) |
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| 摘 要： | 目的:构建靶向NIPSNAP3A的短发夹RNA(shRNA)腺病毒载体,干扰HEK293A细胞中NIPSNAP3A的表达。方法:设计靶向NIPSNAP3A的shRNA,插入穿梭质粒pENTRY,通过Gateway法获得腺病毒颗粒pAd-NIPSNAP3A-shRNA,转染HEK293A细胞,在细胞内包装获得腺病毒。结果:重组腺病毒载体经酶切鉴定正确,制备的病毒感染效率高,能显著抑制NIPSNP3A蛋白的表达。结论:干扰HEK293A细胞NIPSNAP3A表达的shRNA重组腺病毒载体构建成功。
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| 关 键 词： | NIPSNAP3A 短发夹RNA 腺病毒载体 |
Construction of Adenovirus Vector and Virus Packaging of shRNA Against NIPSNAP3A |
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| YU Hua-Ji
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YAN Yong-Hong
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TANG Liu-Jun
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YANG Xiao-Ming
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WANG Xiao-Hui
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WANG Si-Ying.Construction of Adenovirus Vector and Virus Packaging of shRNA Against NIPSNAP3A[J].Letters in Biotechnology,2012,23(4):485-487,492. |
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| Authors: | YU Hua-Ji YAN Yong-Hong TANG Liu-Jun YANG Xiao-Ming WANG Xiao-Hui WANG Si-Ying |
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| Institution: | 1* 1.Department of Pathophysiology,Anhui Medical University,Hefei 230032;2.Department of Beijing Proteome Re search Center,Academy of Military Medical Sciences,Beijing 102206;China |
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| Abstract: | Objective: To construct the recombinant adenovirus vector of short hairpin RNA(shRNA) against NIPSNAP3A and explore the knocking-down expression of NIPSNAP3A in HEK293A cell.Methods: The short hairpin oligonucleotide and complementary strands targeting the consensus sequences of NIPSNAP3A were designed and inserted in the shuttle plasmid pENTRY,then adenovirus particles pAd/PL-DEST-NIPSNAP3A-shRNA through the Gateway method was obtained.The adenovirus particles was transfected into HEK293A cells via lipo some mediation to package and amplify pAd/PL-DEST-NIPSNAP3A-shRNA adenovirus.Results: The adenovirus vectors were verified by enzyme digestion and DNA sequencing,the infection efficiency of constructed adenovirus was high,and could obviously inhibit the expression of NIPSNAP3A.Conclusion: The recombinant adenovirus pAd/ PL-DEST-NIPSNAP3A-shRNA was successfully constructed to knockdown the expression of NIPSNAP3A in 293A cell.This result lays the foundation for further study to investigate the function of NIPSNAP3A. |
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| Keywords: | NIPSNAP3A short hairpin RNA adenovirus vector |
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