Construction of baculovirus expression vector of miRNAs and its expression in insect cells
Shen, Xing Jia
Yu, Xue Li
Wang, Zhan Bin
Cheng, Xiang Chao
- 信息技术－已发表论文 
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 flies to 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 premiR-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 DH10Bac/AcNPV forming bacmid-9a which was transfected 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-9a in ipfected Sf21 insect cells.
CitationMOLECULAR GENETICS MICROBIOLOGY AND VIROLOGY，2012,27（2）：85-90