Smad3基因小干扰RNA的序列设计及其表达载体的构建*

doi:10.3969/j.issn.1672-5069.2010.05.004

Abstract

Abstract: Objective To construct an expression vector with small interfering RNAs of Smad3 in vitro. Methods Three possible small interfering RNAs to Smad3 gene were designed and synthesized，subsequently transfected into rat hepatic stellate cells using Liposome 2000. RT-PCR was used to detect the expression of the Smad3 gene. Then the strands were cloned into pRNAT-U6.1/Neo vector to construct the recombinant plasmids and confirmed by sequencing. Results Both RT-PCR and sequencing assays showed that the recombinant vector pRNAT-SiSmad3 was successfully constructed. Conclusion The successful construction of the recombinant plasmid pRNAT-SiSmad3 will be beneficial to further study in the experiment of Smad3 down-regulation.

Key words: Hepatic fibrosis, Smad3, Interference vector, siRNA, HSCP

WANG Zerong，WANG Jianhao，HU Congli，et al.. Designment and vector construction of siRNA sequences to Smad3 gene in vitro[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2010, 13(5): 328-330.

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Designment and vector construction of siRNA sequences to Smad3 gene in vitro

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