title: Construction of baculovirus expression vector of miRNAs and its expression in insect cells.
abstract: 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 ofmiR-9a. The sequence ofpre-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 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 infected Sf21 insect cells.
authors.affiliation: Animal Science and Technology College, He Nan University of Science and Technology, Luoyang City,471003, Henan Province, PR China. huangyong1979111@126.com
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.forename: Quan
authors.initials: Q
authors.lastname: Zou
authors.forename: Xing Jia
authors.initials: XJ
authors.lastname: Shen
authors.forename: Xue Li
authors.initials: XL
authors.lastname: Yu
authors.forename: Zhan Bin
authors.initials: ZB
authors.lastname: Wang
authors.forename: Xiang Chao
authors.initials: XC
authors.lastname: Cheng
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D012831:Bombyx / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D035683:MicroRNAs / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D035683:MicroRNAs