| 摘要: |
| 为了探讨小干涉RNA(siRNA)对靶基因Smad 7 mRNA表达的阻断作用,采用体外转录方法制备Smad 7基因的小干涉RNAs(siRNAs),用脂质体介导瞬时转染BERP35T-2肺癌细胞系,并采用Northern blot杂交检测靶基因mRNA的表达丰度.结果表明,根据Smad 7编码区序列在体外成功地制备了针对2个不同靶序列的siRNAs;Northern blot杂交显示,在转染siRNA的BERP35T-2细胞中,不管是内源性的还是外源性的,Smad 7 mRNA的表达丰度均明显下降.说明Smad 7基因编码区中542～563 bp及701～722 bp 2个区域均是siRNA作用的有效靶序列,本研究设计并制备的siRNAs能有效抑制Smad 7基因的表达. |
| 关键词: 小干涉RNA Smad 7基因 体外转录 瞬时转染 靶序列 |
| DOI： |
| 分类号: |
| 基金项目:国家重点基础研究发展规划973项目(G1998051207) |
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| Study of RNA interference based on the target sequening of Smad 7 gene |
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| Abstract: |
| This research intended to investigate the suppression of siRNA to the expression of the mRNA of target gene Smad 7.The siRNAs of Smad 7 gene were prepared by in vitro transcription.BERP35T-2 lung cancer cell line was transient transfected with siRNAs and Smad 7 expression plasmid using Lipofectamine 2000 reagent.The expression abundance of target gene mRNA was tested with Northern blot hybridization.According to the coding sequence of Smad 7 gene,we successfully synthesized the siRNAs for two different target sequences in vitro.The result of Northern blot hybridization showed that the expression abundance of both endogenous and exogenous Smad 7 mRNA decreased evidently in BERP35T-2 cells transfected with siRNAs.Both regions of 542-563 bp and 701-722 bp in the coding region of Smad 7 gene are effective target sequences which can be acted by siRNA.The siRNAs designed and synthesized in our research can suppress the expression of Smad 7 gene effectively. |
| Key words: small interfering RNA Smad 7gene in vitro transcription transient transfection target sequences |
