| 栾坤业1 于健1 姚如永2 张炳远1 孙传东1△.TRAIL 慢病毒载体构建及其在肝癌细胞中的表达[J].,2012,12(9): |
| TRAIL 慢病毒载体构建及其在肝癌细胞中的表达 |
| Construction of Lentiviral Vector Carrying TRAIL Gene and Its Expressionin Hepatocarcinoma Cells |
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| DOI： |
| 中文关键词: TRAIL 慢病毒载体 HepG2 |
| 英文关键词: TRAIL Lentiviral vector HepG2 |
| 基金项目:山东省教育厅课题项目基金资助（J11L65） |
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| 中文摘要: |
| 目的：构建携带TRAIL 基因的慢病毒表达载体并实现其在肝癌细胞株HepG2 中的稳定高表达。方法：构建TRAIL 重组慢
病毒表达载体pCDH-CMV-TRAIL-EF1-GFP-T2A-Puro，脂质体法将重组慢病毒载体和包装质粒混合物共转染293T 细胞，包装产
生慢病毒颗粒，纯化并测定病毒滴度。利用Western blotting 检测TRAIL 蛋白在HepG2 中的表达。结果：酶切以及测序证实，成功
构建TRAIL 基因重组慢病毒载体，纯化的慢病毒滴度为1.02×104 ifμ/μL。利用嘌呤霉素筛选获得稳定表达TRAIL 的细胞系，经
Western blot 方法检测到TRAIL 蛋白的稳定高表达。结论：成功构建了带有TRAIL 基因的慢病毒载体，并实现其在HepG2 的稳
定高表达。 |
| 英文摘要: |
| Objective: To construct lentiviral expression vector carrying TRAIL gene and realize its stable high expression in hepatocellular
carcinoma cell line HepG2. Methods: PCDH-CMV-TRAIL-EF1-GFP-T2A-Puro recombinant lentiviral vector was constructed.
The 293T cells were contransfected with the recombinant lentiviral vector and lentivirus package plasmid to produce lentiviral particles
by lipofectamine method. The lentivirus was purified and virus titer was measured. Western blot was used to detect the expressions of
TRAIL protein. Results: The recombinant lentiviral vector carrying TRAIL was confirmed by restriction end nuclease analysis and DNA
sequencing. Virus titer reached to 1.02×104 ifμ/μL. After puromycin selection, the stable high expressions of TRAIL protein were confirmed
by Western blotting. Conclusion: Lentiviral vector carrying TRAIL gene has been successfully constructed realizes its stable high
expression in HepG2 cells. |
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