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出　　处：《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》2013年第12期1131-1131,共1页浙江大学学报（英文版）B辑（生物医学与生物技术）

摘　　要：MicroRNAs （miRNAs） are small endogenous RNAs molecules, approximately 21-23 nucleotides in length, which regulate gene expression by base-pairing with 3＇ untranslated regions （UTRs） of target mRNAs. However, the functions of only a few miRNAs in organisms are known. Recently, the expression vector of artificial miRNA has become a promising tool for gene function studies. Here, a method for easy and rapid construction of eukaryotic miRNA expression vector was described. The cytoplasmic actin 3 （A3） promoter and flanked sequences of miRNA-9a （miR-9a） precursor were amplified from genomic DNA of the silkworm （Bombyx mori） and was inserted into pCDNA3.0 vector to construct a recombinant plasmid. The enhanced green fluorescent protein （EGFP） gene was used as reporter gene. The Bornbyx mori N （BmN） cells were transfected with recombinant miR-9a expression plasmid and were harvested 48 h post transfection. Total RNAs of BmN cells transfected with recombinant vectors were extracted and the expression of miR-9a was evaluated by reverse transcriptase polymerase chain reaction （RT-PCR） and Northern blot. Tests showed that the recom- binant miR-9a vector was successfully constructed and the expression of miR-9a with EGFP was detected.Construction and detection of expression vectors of microRNA-9a in BmN cells Authors:Yong HUANG,Quan ZOU,Sheng-peng WANG,Shun-ming TANG,Guo-zheng ZHANG,Xing-jia SHEN doi:10.1631/jzus.B1000296J.Zhejiang Univ.-Sci.B(Biomed.&Biotechnol.),2011 Vol.12 No.7 P.527-533Abstract:MicroRNAs(miRNAs)are small endogenous RNAs molecules,approximately 21–23 nucleotides in length,which regulate gene expression by base-pairing with 3′untranslated regions(UTRs)of target mRNAs.

关 键 词：小分子RNA 家蚕细胞 表达载体 快速施工 增强型绿色荧光蛋白 检测 逆转录聚合酶链反应 miRNA 

分 类 号：S881[农业科学—特种经济动物饲养]