| 蔡忠良1 阴继凯1 韩腾龙2 张健2 鲁建国1△.用Gateway 系统构建携带Grb2-SH2 基因重组慢病毒载体[J].现代生物医学进展英文版,2012,12(4):616-618. |
| 用Gateway 系统构建携带Grb2-SH2 基因重组慢病毒载体 |
| Construction of Grb2-SH2 genetic restructuring-slowvirus carrier with Gateway system |
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| DOI： |
| 中文关键词: 生长因子受体连接蛋白 重组慢病毒载体 表皮生长因子受体 |
| 英文关键词: Grb2-SH2 Restructuring-slow virus carrier EGFR |
| 基金项目:国家自然青年科学基金资助项目(30901457) |
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| 中文摘要: |
| 目的：构建携带Grb2-SH2 基因重组慢病毒载体。方法: 把Grb2-SH2 基因从合成的工程载体PUC-57 中导到入门载体
pentr-3C 中，再利用LR 反应重组到目的载体plenti,通过酶切和测序分析对比验证Grb2-SH2 基因后，将plenti- Grb2-SH2 质粒利
用慢病毒转染系统转染人胚胎肾上皮细胞株293T 细胞获得携带Grb2-SH2 慢病毒载体。结果：构建的重组质粒经酶切及测序和
分析比对鉴定正确，目的基因片段大小约为500bp；该质粒与包装质粒共转染293T 细胞获取的慢病毒，转染效率约为90%。结
论：成功构建转载质粒plenti- Grb2-SH2 及携带Grb2-SH2 基因慢病毒载体。 |
| 英文摘要: |
| Objective: Construct Grb2-SH2 genetic recombination slow virus carrier. Methods: Put the Grb2-SH2 genes into
introductory carrier pentr-3 C from synthetic engineering carrier PUC-57, reuse LR reaction to recombinate into the purpose plenti
carrier, after comparing and testing gene Grb2-SH2 through the enzyme cut and sequencing analysis and comparison ,use slow virus
carrying system to transfect human embryonic renal epithelial cell lines 293 T cells acquring Grb2-SH2 slow virus carrier. Results: The
restructuring plasmid proved to be correct by the enzyme cut and sequencing analysis and the purpose genetic fragments is about 500 bp;
The efficiency of the plasmid and packaging plasmid transfect 293 T cells get slow is about 90%. Conclusion: Successful construction
reproduced plasmid plenti-Grb2-SH2 and Grb2-SH2 gene slow virus carrier. |
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