Inhibition of GFP expression in hepatocellular carcinoma cells by shRNA

YAN Ge; et al

doi:10.13406/j.cnki.cyxb.2003.06.001

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Inhibition of GFP expression in hepatocellular carcinoma cells by shRNA

更新时间：2024-01-17

- Inhibition of GFP expression in hepatocellular carcinoma cells by shRNA
- Journal of Chongqing Medical University Issue 6, Pages: 681-684(2003)
- 作者机构：
  
  1. 重庆医科大学病毒性肝炎研究所
  2. 重庆医科大学病毒性肝炎研究所,重庆,400010
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2003.06.001
  CLC： R735.7
- Published：2003，
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[1]闫歌,黄爱龙,唐霓,张秉强,向明确,蒲丹.shRNA抑制外源绿色荧光蛋白在肝癌细胞株中的表达[J].重庆医科大学学报,2003(06):681-684.

YAN Ge, et al. Inhibition of GFP expression in hepatocellular carcinoma cells by shRNA. [J]. Journal of Chongqing Medical University (6):681-684(2003)
[1]闫歌,黄爱龙,唐霓,张秉强,向明确,蒲丹.shRNA抑制外源绿色荧光蛋白在肝癌细胞株中的表达[J].重庆医科大学学报,2003(06):681-684. DOI： 10.13406/j.cnki.cyxb.2003.06.001.

YAN Ge, et al. Inhibition of GFP expression in hepatocellular carcinoma cells by shRNA. [J]. Journal of Chongqing Medical University (6):681-684(2003) DOI： 10.13406/j.cnki.cyxb.2003.06.001.

摘要

目的 :构建绿色荧光蛋白（GFP）的短发夹状RNA（shRNA）

观察其在哺乳动物细胞中抑制外源GFP的表达。方法 :设计、合成含GFP编码基因片段

中间被 4个bp间隔的反向重复序列

与转录载体 pTZU6 +1连接

构建pshRNA—GFP重组质粒

与pEGFP -C1共转染肝癌细胞株HepG2和SMMC - 772 1

检测其对GFP表达的影响。结果 :pshRNA—GFP对共转染的 pEGFP -C1中的绿色荧光蛋白有明显的抑制作用

抑制率达 70 %～ 85 %。结论 :构建的 pshRNA—GFP重组质粒能有效地抑制GFP在哺乳动物细胞中的表达。

Abstract

Objective:To construct the plasmid containing short hairpin RNA(shRNA) of green fluorescent protein(GFP) to suppress the expression of GFP.Methods:A 20bp reverse repeated motif of GFP target sequence with 4bp spacer were synthesized

and inserted into pTZU6+1 to generate the pshRNA—GFP plasmid;pshRNA—GFP and pEGFP-C1 or pTZU6+1 and pEGFP-C1 were cotransfected into mammalian cells to detect effect of GFP expression separately.Results:pshRNA—GFP suppressed the GFP expression by 70%～85% in mammalian cells.Conclusion:The result shows that the short hairpin RNA of GFP can efficiently suppress its expression in HepG2 and SMMC-7721.

关键词

RNA干扰绿色荧光蛋白短发夹状RNA

Keywords

RNA interferinggreen fluorescent proteinShort hairpin RNA

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