MDR1、GSTπ特异性siRNA真核表达载体的构建及表达
FUDAN UNIVERSITY JOURNAL OF MEDICAL SCIENCES(2006)
Abstract
目的构建多药耐药基因(MDR1)、谷胱甘肽S-转移酶(GSTπ)特异性小干扰RNA(siRNA)真核表达载体并检测其表达.方法参照siRNA模板设计原则,设计并化学合成2条siRNA模板序列,退火后将其插入质粒pSilencer2.1-U6,限制性酶切和基因测序进行鉴定.脂质体介导下转染K562/Adr细胞,实时荧光定量PCR分析MDR1、GSTπ mRNA的表达,荧光免疫组化检测Pgp、GSTπ蛋白的表达.结果重组质粒pSilencer2.1-MDR1、GSTπ经酶切、测序分析表明siRNA模板序列成功插入预计位点,并且序列正确.用pSilencermdr1、pSilencer2.1-GSTπ分别转染K562/Adr细胞株,mdr1 mRNA表达量下降了71.5%,GSTπ mRNA表达量下降了39.8%,荧光免疫组化显示Pgp、GSTπ表达均显著减少(P<0.01).结论成功构建了MDR1、GSTπ特异性siRNA真核表达载体,该载体可不同程度逆转K562/Adr细胞的多药耐药.
MoreTranslated text
Key words
specific sirna expression vector,mrna,mdr1
AI Read Science
Must-Reading Tree
Example
![](https://originalfileserver.aminer.cn/sys/aminer/pubs/mrt_preview.jpeg)
Generate MRT to find the research sequence of this paper
Chat Paper
Summary is being generated by the instructions you defined