| 应用逆转录病毒载体转染K562细胞株的实验研究 |
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| 引用本文： | 王存邦,白海,薛志文,杨小亮. 应用逆转录病毒载体转染K562细胞株的实验研究[J]. 西北国防医学杂志, 2008, 29(6) |
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| 作者姓名： | 王存邦 白海 薛志文 杨小亮 |
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| 作者单位： | 兰州军区兰州总医院全军血液病中心,甘肃,兰州730050 |
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| 基金项目： | 军队杰出人才基金 ， 兰州军区科研项目 |
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| 摘 要： | 目的:探讨多种细胞因子联合刺激下,逆转录病毒载体对K562细胞的基因转移效率。方法:以PA317-GCGPXSN细胞制备病毒上清,NIH3T3细胞测定病毒滴度后,取经过SCF、IL-3、IL-6预培养刺激后的K562细胞,实施基因转移,流式细胞仪及PCR方法测定基因转移效率。结果:①病毒滴度为1.9×105CFU/m l;②对照组K562细胞测定的荧光强度数值为0.56%,实验组为77.16%,两者差异有统计学意义(P<0.01)。PCR方法扩增到了NeoR基因的特异性片断。结论:细胞因子预刺激后实施基因转移,可有效地将外源基因转移进入K562细胞基因组中。
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| 关 键 词： | K562细胞 细胞因子 逆转录病毒载体 基因转移 绿色荧光蛋白 |
Experimental study on gene transfer into K562 cells mediated by retroviral vector |
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| WANG Cun-bang,BAI Hai,XUE Zhi-wen,et al.. Experimental study on gene transfer into K562 cells mediated by retroviral vector[J]. Medical Journal of National Defending Forces in Northwest China, 2008, 29(6) |
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| Authors: | WANG Cun-bang BAI Hai XUE Zhi-wen et al. |
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| Affiliation: | WANG Cun-bang; BAI Hai; XUE Zhi-wen; et al.(Department of Hematology; Lanzhou General Hospital; Lanzhou Command; PLA; Lanzhou 730050; China); |
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| Abstract: | Objective:To explore gene transfer efficiency on K562 cells mediated by retroviral vector and stimulated with cytokines.Methods: The viral supernatants were harvested with PA317-GCGPXSN and detected with NIH3T3 cells.Then gene transfer was made using K562 cells as target cells which were stimulated by SCF,IL-3 and IL-6.The gene transfer efficiency was detected by FACS and PCR.Results:The viral titer was 1.9×105 CFU/ml;The gene transfer efficiency of experiment group was 77.16%,whereas the control group was ... |
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| Keywords: | K562 cell Cytokine Retroviral vector Gene transfer Green fluorescent protein |
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