摘要
目的构建单核细胞趋化蛋白-1(MCP-1)小分子干扰RNA(siRNA)表达载体,为进一步研究其对动脉粥样硬化(AS)的防治作用提供手段。方法设计并合成两端含有酶切位点两条DNA序列,经退火成互补双链,再克隆至载体pSi-lencer 2.0-U6中构建重组表达载体,转化DH-5α菌株,提取质粒行双酶切鉴定,并进行序列测定。结果双酶切证实MCP-1 siRNA表达载体克隆构建成功,插入片段测序结果与合成的siRNA结果一致。结论成功构建MCP-siRNA表达载体,为动脉粥样⒉化的防治奠定实验基础。
Objective To construct the small interfering RNA（siRNA） expression vector for monocyte chemotactite protein-1（MCP-1） and provide ways for artery stenosis treatment. Methods Two DNA sequences containing the sites of restriction endonuclease at both ends were designed and synthesized. The eomplement form was obtained by annealing and being cloned into vector pSilencer2.0-U6 and the recombinant plasmid was transformed into strain DH-5 α. The plasmid identified by restriction enzyme was used for sequencing. Results MCP-1 siRNA expression vector was successfully constructed and identified by double endonuclease digestion. Sequence analysis of the inserted fragment revealed the salne sequence as that of the synthesized siRNA oligonucleotides. Conclusion MCP-1 siRNA expression vector has been successfully constructed, which lays the basis for its application in the prevention and treatment of artery stenosis.
出处
《中华普通外科学文献（电子版）》
2008年第5期10-12,共3页
Chinese Archives of General Surgery(Electronic Edition)
基金
广东省科技计划重点引导项目资助(2005B31201001)
教育部高等学校博士点专项基金项目资助(20050558053)
