| 稳定敲低MYH10基因细胞株的建立 |
| 投稿时间：2015-03-29 修订日期：2015-04-15 点此下载全文 |
| 引用本文：周晨辰,陆琤,张鹏飞,张硌.稳定敲低MYH10基因细胞株的建立[J].医学研究杂志,2015,44(11):51-53 |
| DOI：
10.11969/j.issn.1673-548X.2015.11.014 |
| 摘要点击次数: 1354 |
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| 基金项目:国家自然科学基金青年科学基金资助项目(31400739);北京市自然科学基金资助项目(5144033) |
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| 中文摘要:目的 构建稳定敲低MYH10基因的COS-7细胞株。方法 在293TX细胞中进行病毒的包装,用获得的高效价慢病毒感染COS-7细胞,通过免疫印迹和荧光显微镜,检测病毒感染后COS-7细胞MYH10蛋白表达的变化。结果 在构建的两种重组质粒中,MYH10-lentiviral shRNA-2在转染后表现出明显的干扰作用,感染后能明显下调COS-7细胞的MYH10蛋白表达。结论 病毒感染的COS-7细胞可明显抑制COS-7细胞内源MYH10的表达,说明了稳定敲低MYH10基因的COS-7细胞株的建立,为进一步研究MYH10在相关疾病中的功能奠定了基础。 |
| 中文关键词:MYH10基因 COS-7细胞株 慢病毒感染 |
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| Establishment of Stable COS-7 Cell Strain with MYH10-knock-down. |
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| Abstract:Objective To establish COS-7 cell strain stable knockdown of MYH10 gene. Methods The lentiviral plasmids were transfected into 293TX cells to package into lentiviral particles. Then COS-7 cells was transduced with the lentiviral particles. Western blot and fluorescence microscope were carried out to analyze the suppression of MYH10 in COS-7 Cells. Results In the construction of two recombinant plasmid, MYH10-lentiviral shRNA-2 showed significant interference after transfection, and can obviously reduced expression of MYH10 in COS-7 cells. Conclusion Infected COS-7 cells can inhibit the expression of the MYH10 COS-7 cells, indicating that a stable COS-7 cell strain has been successively established. |
| keywords:MYH10 gene COS-7 cell strain Lentivirus transfection |
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