Identification Of Bmyan, A Target Gene Of Bmo-miR-7

MiR-7 and yan gene ragulate the differentiation of photoreceptor and development of complex eyes in Drosophila melanogaster.As the representative of Lepidoptera. bombyx mori also has a pair of complex eyes, but it still remains unknown whether miR-7 and van are collectively regulating the complex eyes of this animal. Here we indentified yan gene in silkworm and comfirmed one target site of bmo-miR-7 on its 3'UTR.1. Cloning and expression pattern analysis of BmyanBmyan consists of 4 exons and 3 introns and encodes 477 amino acid residues. SAM-PNT and a ETs domains were found in the protein. The former domain widely exists in signal transduction pathways while the latter regulates transcription of specific genes.RT-PCR revealed high expression of Bmyan in the head, ovary of day-3 fifth instar silkworm, low expression in the body wall and midgut. and no accumulation in testis. hemocyte and other organs. Furthermore, it was stably expressed in the larval and pupal stages.2.3'RACE and sequence analysis of BmyanThe 3'UTR of Bmyan is 568bp, which contains a potential mir-7 recognition site locates from 98th to 116th nucleotide residues.3. Target validation of Bmyan via dual-luciferase assay.Dual-luciferase assay was performed to reveal the possible target site of bmo-miR-7. bmo-miR-7 and luciferase-UTR report vector were co-transfected into BmE cell line, resulting in lower luciferase activity than that from the negative control. Accordingly. Bmyan was a target gene of bmo-miR-7.