真核表达Hsa-miR-1基因及其检测方法的建立  

Eukaryotic Expression of Hsa-miR-1 Gene and Establishment of Its Detection Method

作　　者：胡徐庞[1] 李伟[1] 曹跃芬[2] 张璇[1] 黄青红[1] 徐科[1] 杨耿兵[1] 魏旭斌[1] 钱程[1] 刘立[1] 

机构地区：[1]浙江理工大学生命科学学院新元医药研究所,杭州310018 [2]浙江理工大学生物工程研究所,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2011年第5期783-788,共6页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：国家重点基础发展计划资助项目(973)(2010CB529406)

摘　　要：获得Hsa-miR-1的方法主要由人工合成,需耗费很高的成本与时间。文章通过将Hsa-miR-1前体部分连接至pcDNA3.0,可实现Hsa-miR-1基因的高表达。构建含有Hsa-miR-1基因表达前体的表达载体pcDNA-Hsa-miR-1和含有与Hsa-miR-1基因完全互补序列的检测载体pEGFP-C1-CM1。通过共转染pcDNA-Hsa-miR-1和pEGFP-C1-CM1检测Hsa-miR-1基因的表达,并采用Taqman探针法定量检测Hsa-miR-1基因表达水平,成功构建了可高效表达Hsa-miR-1基因的真核生物质粒pcDNA-Hsa-miR-1,并首次建立起Hsa-miR-1基因的检测系统pEGFP-C1-CM1,该检测系统的建立将为今后Hsa-miR-1在肿瘤细胞中的研究提供准确、快捷的检测方法。Currently available Hsa-miR-1 mainly by the synthetic methods,is very costly and time-consuming.By part precursor of the Hsa-miR-1 connecting to the pcDNA3.0,can be realized Hsa-miR-1 genes were highly expressed.Constructing containing Hsa-miR-1 precursor gene expression vector pcDNA-Hsa-miR-1 and containing Hsa-miR-1 gene completely complementary sequence detection vector pEGFP-C1-CM1,and using Taqman probes for quantification of Hsa-miR-1 gene expression level,can successfully constructe and highly expressed Hsa-miR-1 genes in eukaryotic plasmid pcDNA-Hsa-miR-1.It is the first time to establish Hsa-miR-1 gene detection system pEGFP-C1-CM1,and the detection system for Hsa-miR-1 in tumor cells study can provide accurate and quick detection method in future.

关 键 词：Hsa-miR-1载体 Taqman探针检测 pcDNA-Hsa-miR-1 pEGFP-C1-CM1 

分 类 号：Q786[生物学—分子生物学]