| 人CD59 RNAi逆转录病毒载体系统的构建与鉴定 |
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| 引用本文： | 石学香,高美华,臧金林.人CD59 RNAi逆转录病毒载体系统的构建与鉴定[J].青岛大学医学院学报,2009,45(1). |
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| 作者姓名： | 石学香 高美华 臧金林 |
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| 作者单位： | 1. 青岛大学医学院免疫学教研室,山东,青岛,266071
2. 青岛市市立医院 |
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| 摘 要： | 目的 利用siRNA表达载体法构建并筛选携带针对CD59基因pSUPER retro RNAi逆转录病毒载体及稳定产毒的细胞克隆.方法 用DNA重组技术,将3条60 bp能转录产生靶向CD59小发夹RNA(shRNA)的寡核苷酸序列定向克隆入逆转录病毒载体pSUPER retro,脂质体法转染包装细胞系Phoenix A,建立产生逆转录病毒的细胞克隆.结果 重组载体经PCR及限制性内切酶酶切鉴定初步成功后测序序列正确;重组载体转染包装细胞,可表达绿色荧光蛋白,表明包装成功.结论 特异性沉默CD59基因的pSUPER retro RNAi逆转录病毒载体以及稳定产毒的细胞系构建成功,为后续进行肿瘤细胞的研究奠定了基础,可望为肿瘤治疗开辟新途径.
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| 关 键 词： | RNA 双链 抗原 CD59 转染 逆转录病毒载体 |
CONSTRUCTION AND IDENTIFICATION OF RETRO RNAi SYSTEM OF THE SPECIFIC SILENCING CD59 |
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| SHI XUE-XIANG,GAO MEI-HUA,ZANG JIN-LIN.CONSTRUCTION AND IDENTIFICATION OF RETRO RNAi SYSTEM OF THE SPECIFIC SILENCING CD59[J].Acta Academiae Medicinae Qingdao Universitatis,2009,45(1). |
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| Authors: | SHI XUE-XIANG GAO MEI-HUA ZANG JIN-LIN |
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| Institution: | SHI XUE-XIANG,GAO MEI-HUA,ZANG JIN-LIN (Department of Immunology,Qingdao University Medical College,Qingdao 266071,China) |
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| Abstract: | Objective To construct recombinant retroviral vectors that target CD59 gene and a stable virus-producing cell line.Methods Three 60 bp encoded targeting CD59 gene shRNA sequences were cloned into pSUPER with DNA recombinant technique.The packaging cell Phoenix A was transfected with this recombinant plasmids using liposome.Results The result of electrophoresis of PCR,enzyme digestion and sequence demonstrated that 60 bp had been inserted into the vector.And virus-producing cell lines were selected.Conclusio... |
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| Keywords: | RNA double-stranded Antigens CD59 Transfection Retroviral vector |
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