| NY-ESO-1抗原T细胞受体基因慢病毒载体的构建及鉴定 |
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| 引用本文： | 顾娜,徐珩,冯丹丹,李宁,闾军. NY-ESO-1抗原T细胞受体基因慢病毒载体的构建及鉴定[J]. 北京医学, 2010, 32(12): 951-954 |
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| 作者姓名： | 顾娜 徐珩 冯丹丹 李宁 闾军 |
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| 作者单位： | 首都医科大学附属北京佑安医院肝病消化科,100069;安徽医科大学 |
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| 基金项目： | 北京市自然科学基金，留学人员科技活动项目择优资助 |
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| 摘 要： | 目的构建NY-ESO-1抗原T细胞受体基因（TCellReceptor,TCR）的慢病毒载体并获得重组慢病毒。方法设计特异性NY-ESO-1TCR的引物,用聚合酶链反应（PCR）对我们前期已克隆的NY-ESO-1TCRcDNA进行体外扩增,双酶切后定向插入到pCL20c-MSCV-GFP质粒,构建NY-ESO-1抗原特异性TCR基因的重组慢病毒载体pCL20c-MSCV-ESOTCR,并经酶切、PCR及测序鉴定。用pCL20c-MSCV-ESOTCR﹑pCL20c-HIV-gp、pCAG4-RTR2和CAG-VSV-G共转染T293包装细胞,获得上清,上清浓缩后感染Hela细胞,观察ESOTCR基因及蛋白水平的表达。结果经PCR及测序证实,成功构建重组慢病毒载体pCL20c-MSCV-ESOTCR,包装产毒后感染Hela细胞,RT-PCR法及荧光显微镜均能检测及观察到基因及蛋白水平的表达。结论成功构建了表达NY-ESO-1抗原T细胞受体基因（TCellReceptor,TCR）的重组慢病毒载体。
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| 关 键 词： | NY-ESO-1T细胞受体 慢病毒载体 Hela细胞 |
Construction and expression in Hela cell of NY-ESO-1 antigen T cell receptor lentiviral vector |
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| GU Na,XU Heng,FENG Dan-dan,et al. Construction and expression in Hela cell of NY-ESO-1 antigen T cell receptor lentiviral vector[J]. Beijing Medical Journal, 2010, 32(12): 951-954 |
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| Authors: | GU Na XU Heng FENG Dan-dan et al |
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| Affiliation: | GU Na,XU Heng,FENG Dan-dan,et al(Department of Hepatology and Gastroenterology,Beijing You'an Hospital,Beijing 100069) |
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| Abstract: | Objective To establish an NY-ESO-1(New York esophageal squamous cell carcinoma 1) T Cell receptor(TCR) recombinant lentiviral vector.Methods Specific NY-ESO-1 TCR primers were designed,and NY-ESO-1 TCR gene was amplified by PCR(polymerase chain reaction) with the template of NY-ESO-1 TCR cDNA,which we had cloned before in our lab.The PCR products were digested by EcoR I and Not I,and directly cloned into pCL20c-MSCV-GFP vector named pCL20c-MSCV-ESOTCR and confirmed by digestion,PCR and sequencing.pCL20c-MSC... |
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| Keywords: | NY-ESO-1 TCR lentiviral vector Hela cell |
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