CONSTRUCTION OF BACULOVIRUS EXPRESSION VECTOR OF miRNAs AND ITS EXPRESSION IN INSECT CELLS

文献类型： 外文期刊

第一作者： Huang, Yong

作者： Huang, Yong;Yu, Xue Li;Wang, Zhan Bin;Cheng, Xiang Chao;Huang, Yong;Shen, Xing Jia;Zou, Quan

作者机构：

关键词： Baculovirus expression system;MiR-9a;AcNPV;Sf21 cells

期刊名称：MOLECULAR GENETICS MICROBIOLOGY AND VIROLOGY （ 影响因子：0.352； 五年影响因子：0.526 ）

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收录情况： SCI

摘要： MicroRNAs (miRNAs) are endogenous small non-protein coding RNAs that play important regulatory roles in animals and plants by binding to target transcripts for cleavage or translational repression. The miR-9a is very conservative in animals from fliesto humans. Studies indicated that miR-9a is involved in the regulation of neurogenesis in animals. In our study, the baculovirus expression system was used to transcribe a recombinant vector containing miR-9a for further analysis the function of miR-9a.The sequence of pre-miR-9a from silkworm DNA was first cloned into the donor pFastBac. The enhanced green fluorescent protein (EGFP) was used as reporter gene. The recombinant donor plasmid pFastBac-miR-9a was transformed into E.coli DHlOBac /AcNPV forming Bacmid-9a which was trans-fected into insect cells with cational lipofectin. The transcription of mature miR-9a was detected by Real-time PCR. The results show the recombinant Bacmid-9a was successfully constructed and effectively transcribed miR-9ain infected Sf21 insect cells.

分类号： Q75

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