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脂质体介导外源基因体外转染牛胎儿成纤维细胞条件的优化 被引量:23

Optimization of Parameters of Exogene Transfection ofBovine Fetal Fibroblasts in vitro Mediated by Liposome
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摘要 通过脂质体(FuGENE-6)介导,将真核表达载体pEGFP-C1成功导入体外培养的牛胎儿成纤维细胞,探讨影响外源基因转染效率的参数,如DNA和脂质体的用量、转染的细胞数量以及细胞暴露于DNA与脂质体复合物的时间长度。通过实验发现,绿色荧光蛋白(greenfluorescentprotein,GFP)基因的表达随DNA、脂质体量的增加而增加,延长细胞暴露时间反而使转染效率下降,转染细胞数适当才能得到较高的转化率。 pEGFP-C1 eucaryon expression vector was successfully transfected by liposome into bovine fetal fibroblasts.We investigated the effect of parameter such as the dose of DNA and liposome,number of cell transfected and exposure time of the cell to the DNA-liposome complexes.It was indicated that GFP(green fluorescent protein)expression was enhanced as the dose of DNA and liposome increased and on decline as the exposure time was prolonged.The improvement of transfection efficiency depent on the suitable cell number.
出处 《遗传》 CAS CSCD 北大核心 2002年第6期653-655,共3页 Hereditas(Beijing)
基金 内蒙古大学优秀青年教师培养基金(NMU-200102)资助
关键词 脂质体介导 外源基因体外转染 牛胎儿成纤维细胞 优化 绿色荧光蛋白 转染条件 liposome GFP bovine fetal fibroblasts transfection
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参考文献3

  • 1[1]Felgner P L,et al.Liposome:a hig1`hly efficiency lipid-mediated DNA transfection procedures[J].Proc Natl Acad Sci USA,1987,84 :7413~7417.
  • 2[2]Stamatutos L,et al.Interactions of cationic lipid vesicles with negatively charged phospholipid vesicles and biology membranes[J].Biochemistry,1988,27:3917~3925.
  • 3[3]Sambrook J,et al.Molecular Cloning:A laboratory manual,2 nd Ed[M].Cold Spring Harbor Laboratory Press,Cold Spring Harbor,NY,1989.

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