影响生长抑素重组质粒pEGS/2SS转染HeLa细胞效果的因素研究

畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (3): 279-285.doi:

影响生长抑素重组质粒pEGS/2SS转染HeLa细胞效果的因素研究

何晓红1,4，曹少先2，茆达干1，杨利国3*

1.南京农业大学动物科技学院，南京210095；2. 江苏省农业科学院畜牧研究所，南京 210014；3.华中农业大学动物科技学院，武汉 430070; 4.中国农业科学院北京畜牧兽医研究所,北京 100193

收稿日期:2008-11-25 修回日期:1900-01-01 出版日期:2010-03-24 发布日期:2010-03-24
通讯作者: 杨利国

Transient Transfection Factors for Expression of Recombinant Somatostatin Plasmid pEGS/2SS in Cultured HeLa Cells

HE Xiaohong ^1,4, CAO Shaoxian ², MAO Dagan¹, YANG Liguo ³*

1.College of Animal Science and Technology, Nanjing Agricultural University,NanJing 210095, China; 2.Institute of Animal Science, Jiangsu Academy of Agricultural Sciences,Nanjing 210014, China; 3.College of Animal Science and Technology,Huazhong Agricultural University, Wuhan 430070, China； 4.Institute of Animal Science,Chinese Academy of Agricultural Sciences, Beijing 100193,China

Received:2008-11-25 Revised:1900-01-01 Online:2010-03-24 Published:2010-03-24
Contact: YANG Liguo

摘要/Abstract

摘要： 旨在分析生长抑素重组质粒pEGS/2SS转染HeLa细胞（人宫颈癌上皮细胞）转染效果的影响因素，探讨最佳转染条件，为进一步研究生长抑素基因疫苗的作用机制和作用效果奠定基础。以脂质体转染法将带有绿色荧光蛋白（GFP）报告基因的重组质粒pEGS/2SS转染HeLa细胞，探讨质粒转染HeLa细胞最佳条件的4个参数：质粒DNA剂量、脂质体剂量、最佳转染时间和质粒提取方法，在荧光显微镜下观察细胞转染情况并计算转染率。结果：在六孔细胞培养板上，当DNA/脂质体剂量为1 μg/6 μg时，转染效率最高，在转染后72 h达到34.02%；当用Plasmid Maxi Kit试剂盒提取质粒，转染时间为48 h，转染效率最高，达到17.88%。重组质粒pEGS/2SS转染HeLa细胞条件是：采用试剂盒（Plasmid Maxi Kit）提取的质粒，DNA和脂质体的剂量分别为1 μg与6 μg，转染时间48 h，此时的转染效率最高，达17.88%。

Abstract: We aimed to investigate the factors infecting transfection efficiency of somatostatin recombinant plasmid pEGS/2SS introducting into HeLa cells, make preparations for advanced study on somatostatin gene vaccine mechanism and effect of action. Four parameters including transfection reagent (lipofectamineTM) concentration, DNA concentration, cells cultured time after transfection and DNA extracting techniques were analyzed and optimized, and transfection efficiency was evaluated by the percentage of transfected cells with green fluorescence under fluorescence microscope. Based on the optimized lipofectamineTM and DNA concentration, 34.02% transfected HeLa cells could express green fluorescence 72 h after transfection when the concentration of lipofectamineTM and DNA was individually 6 μg and 1 μg; Based on the optimized cells cultured time after transfection and DNA extracting techniques, 17.88% transfected HeLa cells could express green fluorescence protein with Plasmid Maxi Kit extracting DNA 48 h after trasfection. Optimal transfection conditions were determined with four parameters, 17.88% HeLa cells could express green fluorescence protein with Plasmid Maxi Kit extracting DNA, 1 μg DNA and 6 μg lipofectamineTM 48 h after transfection.

何晓红;曹少先;茆达干;杨利国. 影响生长抑素重组质粒pEGS/2SS转染HeLa细胞效果的因素研究[J]. 畜牧兽医学报, 2010, 41(3): 279-285.

HE Xiaohong;CAO Shaoxian;MAO Dagan;YANG Liguo . Transient Transfection Factors for Expression of Recombinant Somatostatin Plasmid pEGS/2SS in Cultured HeLa Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2010, 41(3): 279-285.

0
/ / 推荐

导出引用管理器 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

链接本文: https://www.xmsyxb.com/CN/

https://www.xmsyxb.com/CN/Y2010/V41/I3/279