摘要
目的考察并确定多聚乙烯亚胺(PEI)最佳的体外细胞转染条件,评价PEI作为基因转染试剂的可行性。方法以MTT法检测不同相对分子质量PEI的细胞毒性。以PEI为载体,在不同的N/P比值条件下,将绿色荧光蛋白基因导入人Ⅱ型肺上皮A549细胞,以荧光显微镜及荧光分光光计系统检测其转染效率。结果相对分子质量为25×103的PEI,当N/P比值为10时,其转染效率最高。结论PEI是一种价格低廉、低毒性、高效率的基因转染载体,可应用于体内外基因治疗。
Abstract
OBJECTIVE To investigate the optimum condition of polyethylenimine(PEI) for gene transfer to A549 cells.METHODS The cytotoxicity of PEI was evaluated by mononuclear cell cytotoxicity assay(MTT).In different conditions, PEI was the vector, and A549 cells were transfected with fluorescent protein(pEGFP-C3) gene.Expression efficiency of the transduced gene was monitored by fluorescence miceoscopy and fluorescence assay system.RESULTS The optimal transfection efficiency of PEI Mw (0.25×103) was obtained at N/P ratio equaled to 10. CONCLUSION PEI is an inexpensive,low cytotoxicity and high efficiency vector.It can be widely used in the area of gene therapy.
关键词
非病毒载体 /
多聚乙烯亚胺 /
基因转染 /
绿色荧光蛋白真核表达质粒 /
人Ⅱ型肺上皮A549细胞
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Key words
non-viral vector /
polyethylenimine /
transfection efficiency /
pEGFP-C3 /
A549
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赵梦丹;杨峰亮;陈坚;虞和永.
多聚乙烯亚胺介导基因转染的体外研究[J]. 中国药学杂志, 2008, 43(16): 1211-1213
ZHO Meng-dn;YNG Feng-ling;CHEN Jin;YU He-yong.
Study on Gene Transfer of Polyethylenimine as a Nonviral Vector in Vitro[J]. Chinese Pharmaceutical Journal, 2008, 43(16): 1211-1213
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参考文献
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脚注
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