bmo-miR-275 down-regulates expression of Bombyx mori sericin gene 2 in vitro

PLoS One. 2018 Jan 30;13(1):e0190464. doi: 10.1371/journal.pone.0190464. eCollection 2018.

Abstract

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, BmN cells were harvested and luciferase activity was detected. Results showed that luciferase activity was reduced significantly (P<0.05) in BmN 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3' Untranslated Regions
  • Animals
  • Bombyx / genetics*
  • Bombyx / growth & development
  • Cell Line
  • Down-Regulation*
  • Genes, Insect*
  • In Vitro Techniques
  • Larva / metabolism
  • MicroRNAs / genetics
  • Sericins / genetics*

Substances

  • 3' Untranslated Regions
  • MicroRNAs
  • Sericins

Grants and funding

This work is funded by the National Nature Science Foundation of China (Grant Nos. 31402143 to PQ, 31672490 to XS), the Nature Science Foundation of JiangSu Province (Grant No. BK20141285 to XS), and the Doctoral Scientific Research Foundation of Jiangsu University of Science and Technology (Grant No. 1732931702 to PQ). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.