Open Accessbmo-miR-275 down-regulates expression of Bombyx mori sericin gene 2 in vitro
Author(s) -
Ping Qian,
Tao Jiang,
Xin Wang,
Fei Song,
Chen Chen,
Xingjia Shen
Publication year - 2018
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0190464
Subject(s) - bombyx mori , sericin , luciferase , microbiology and biotechnology , transfection , reporter gene , untranslated region , plasmid , gene expression , biology , in vitro , three prime untranslated region , microrna , messenger rna , gene , regulation of gene expression , genetics , silk , materials science , composite material
We hypothesized that bmo-miR-275 has a potential regulatory function regarding the expression of sericin gene 2 ( BmSer-2 ). First, we examined the expression of bmo-miR-275 and its target gene BmSer-2 in seven different tissues from 5th instar day-3 silkworm larvae. The results showed that they were both specifically expressed in the middle silk gland, implying that spatio-temporal conditions are required for bmo-miR-275 to regulate the expression of BmSer-2 . To test this hypothesis, we constructed a pri-bmo-miR-275 expressing plasmid pcDNA3.0 [ ie1-egfp -pri-bmo-miR-275-SV40] and BmSer-2 -3´UTR recombinant reporter plasmids pGL3.0 [ A3-luc-Ser-2 -3′ UTR-SV40]. Finally, Bm N cells were harvested and luciferase activity was detected. Results showed that luciferase activity was reduced significantly (P<0.05) in Bm N cells co-transfected with pcDNA3.0 [ ie1-egfp -pri-bmo-miR-275-SV40] and pGL3.0 [ A3-luc-Ser-2 -3’UTR-SV40], suggesting that bmo-miR-275 can down-regulate the expression of BmSer-2 in vitro. Our results improve the understanding of the regulatory function of Bombyx mori miRNA on the expression of genes regulating silk formation.