Based on bioinformatic analysis, we selected two novel micro RNAs（miRNAs）, bmo-miR-0001 and bmomiR-0015, from high-throughput sequencing of the Bombyx mori larval posterior silk gland（PSG）. Firstly, we examined th...Based on bioinformatic analysis, we selected two novel micro RNAs（miRNAs）, bmo-miR-0001 and bmomiR-0015, from high-throughput sequencing of the Bombyx mori larval posterior silk gland（PSG）. Firstly, we examined the expression of bmo-miR-0001 and bmo-miR-0015 in 12 different tissues of the 5th instar Day-3 larvae of the silkworm. The results showed that the expression levels of both bmo-miR-0001 and bmo-miR-0015 were obviously higher in the PSG than in other tissues, implying there is a spatio-temporal condition for bmo-miR-0001 and bmo-miR-0015 to regulate the expression of Bm Fib-L. To test this hypothesis, we constructed pri-bmo-miR-0001 expressing the plasmid pc DNA3.0 [ie1-egfp-pri-bmo-miR-0001-SV40] and pri-bmo-miR-0015 expressing the plasmid pc DNA3.0 [ie1-egfp-pribmo-miR-0015-SV40]. Finally, the Bm N cells were harvested and luciferase activity was detected. The results showed that luciferase activity was reduced significantly（P〈0.05） in Bm N cells co-transfected by pc DNA3.0 [ie1-egfp-pri-bmomiR-0001-SV40] or pc DNA3.0 [ie1-egfp-pri-bmo-miR-0015-SV40] with p GL3.0 [A3-luc-Fib-L-3＇UTR-SV40], suggesting that both bmo-miR-0001 and bmo-miR-0015 can down-regulate the expression of Bm Fib-L in vitro.展开更多
基金supported by the National Natural Science Foundation of China(Nos.31172266/C1703 and 31402143/C1703)
文摘Based on bioinformatic analysis, we selected two novel micro RNAs（miRNAs）, bmo-miR-0001 and bmomiR-0015, from high-throughput sequencing of the Bombyx mori larval posterior silk gland（PSG）. Firstly, we examined the expression of bmo-miR-0001 and bmo-miR-0015 in 12 different tissues of the 5th instar Day-3 larvae of the silkworm. The results showed that the expression levels of both bmo-miR-0001 and bmo-miR-0015 were obviously higher in the PSG than in other tissues, implying there is a spatio-temporal condition for bmo-miR-0001 and bmo-miR-0015 to regulate the expression of Bm Fib-L. To test this hypothesis, we constructed pri-bmo-miR-0001 expressing the plasmid pc DNA3.0 [ie1-egfp-pri-bmo-miR-0001-SV40] and pri-bmo-miR-0015 expressing the plasmid pc DNA3.0 [ie1-egfp-pribmo-miR-0015-SV40]. Finally, the Bm N cells were harvested and luciferase activity was detected. The results showed that luciferase activity was reduced significantly（P〈0.05） in Bm N cells co-transfected by pc DNA3.0 [ie1-egfp-pri-bmomiR-0001-SV40] or pc DNA3.0 [ie1-egfp-pri-bmo-miR-0015-SV40] with p GL3.0 [A3-luc-Fib-L-3＇UTR-SV40], suggesting that both bmo-miR-0001 and bmo-miR-0015 can down-regulate the expression of Bm Fib-L in vitro.
