| iRhom2及其突变基因重组慢病毒的包装和稳定表达细胞系的建立 |
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| 引用本文： | 游颖,马雨楠,曾林,孙兆增.iRhom2及其突变基因重组慢病毒的包装和稳定表达细胞系的建立[J].实验动物与比较医学,2015,23(6):597-601. |
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| 作者姓名： | 游颖 马雨楠 曾林 孙兆增 |
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| 作者单位： | 军事医学科学院实验动物中心,军事医学科学院实验动物中心,军事医学科学院实验动物中心,军事医学科学院实验动物中心 |
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| 基金项目： | 国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| 摘 要： | 目的：通过重组慢病毒感染，建立iRhom2及其突变基因的Vero细胞稳定细胞表达系，用于iRhom2的功能研究。方法：将iRhom2 及其突变基因克隆到慢病毒载体Lenti-OE-Flag，构建重组慢病毒载体Lenti-OE-iRhom2和Lenti-OE-iRhom2mut，将重组质粒瞬时转染HEK-293T包装细胞，获得重组慢病毒。将重组病毒感染Vero细胞，利用puromycin进行加压筛选，获得二者的重组表达细胞系。 结果：构建了iRhom2及其突变基因的逆转录病毒载体，并获得了重组逆转录病毒病毒，并利用该病毒获得了二者的稳定表达细胞系Western-blot实验证实，二者能够在Vero细胞内稳定表达。结论：利用重组逆转录病毒感染，成功获得了iRhom2及其突变系的Vero细胞稳定表达株，为进一步研究iRhom2的生物学功能及其机制奠定了良好的基础。
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| 关 键 词： | iRhom2 慢病毒载体 包装细胞 稳定表达 |
| 收稿时间： | 2015/6/23 0:00:00 |
| 修稿时间： | 2015/7/24 0:00:00 |
Packaging of the recombinant lentivirus of iRhom2 and its mutant and establishment of a stable iRhom2-expressing cell line |
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| YOU Ying,MA Yu-nan,ZENG Lin and SUN Zhao-zeng.Packaging of the recombinant lentivirus of iRhom2 and its mutant and establishment of a stable iRhom2-expressing cell line[J].Laboratory Animal and Comparative Medicine,2015,23(6):597-601. |
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| Authors: | YOU Ying MA Yu-nan ZENG Lin and SUN Zhao-zeng |
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| Affiliation: | Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071, China;Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071, China;Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071, China;Laboratory Animal Center of the Academy of Military Medical Sciences, Beijing 100071, China |
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| Abstract: | Objective:To establish stable trasfection cell lines of iRhom2 and its mutant through the recombinant lentivirus infection . Methods: The full-length gene of iRhom2 its mutant were cloned into the lentivirus vector Lenti-OE-Flag, and got recombinant lentiviral vector of Lenti-OE-iRhom2 and Lenti-OE-iRhom2mut. The constructed recombinant lentivirus vectors were transfected into HEK-293T packaging cells to obtain the recombinant birus. The Vero cells were infected with recombinant virus. The stable expression cell lines were resistance screening Usued puromycin. Results: The recombinant lentivirus vectors were accomplished and packaged the recombinant virus. The stable expression cell lines were obtained using virus infection and the expression of the proteins were testified with Western-blot. Conclusion: The stable expression cell lines of iRhom2 and its mutant were achieved by recombinant lentivirus infection,which paves the way for future study on biological functions and mechanism of iRhom2 |
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| Keywords: | iRhom2 Lentivirus vetor Packaging cell Vero cell line |
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