摘要
miRNAs可通过靶向目标基因3′UTR,负调控目的基因的表达。本研究以马氏珠母贝(Pinctada fucata martensii)的一种miRNA Pfm-miR-9b-5p成熟序列及其基因组数据为基础,获得Pfm-miR-9b-5p前体序列,其长度为99 bp,包含典型的发夹结构。多序列比对结果表明,Pfm-miR-9b-5p的成熟序列和前体序列均与其他物种高度保守。系统进化树分析结果表明,Pfm-miR-9b-5p的前体序列与软体动物聚为一支,与帽贝(Lottia gigantean)亲缘关系最近。组织差异表达分析表明,Pfm-miR-9b-5p在矿化器官边缘膜和套膜区高表达。靶标预测结果表明,贝壳基质蛋白基因PNU9为Pfm-miR-9b-5p的潜在靶标基因之一。注射Pfm-miR-9b-5p mimics过表达Pfm-miR-9b-5p后,靶标基因PNU9的表达被显著抑制,并且贝壳珍珠层和棱柱层均出现异常生长。综上,在马氏珠母贝中,Pfm-miR-9b-5p可通过负向调控矿化基因PNU9的表达,参与贝壳的形成。本研究为进一步探讨miRNAs参与贝壳形成的机制提供了参考。
miRNAs act as the negative regulator involved in shell formation by targeting to the 3′ UTR of the target gene. In this study, we obtained the precursor sequence of Pfm-miR-9b-5p based on the miRNA mature sequence of Pinctada fucata martensii identified before and its genome data. The precursor Pfm-miR-9b-5p had a length of 99 bp, which containing a characteristic hairpin structure. Sequence alignment showed that both the mature sequence and the precursor were highly conserved with other species. Phylogenetic analysis indicated that the precursor sequence of Pfm-miR-9b-5p clustered with mollusk and highly related to Lottia gigantean. Differential expression analysis in tissues showed that Pfm-miR-9b-5p highly expressed in mantle edge and mantle pallial which is mainly responsible for shell formation. The prediction of target genes of Pfm-miR-9b-5p showed that shell matrix protein PNU9 is one of the potential target genes of Pfm-miR-9b-5p. When Pfm-miR-9b-5p was overexpressed by injection of mimics, the expression of target gene PNU9 was significantly suppressed and the nacre layer and prismatic layer grew abnormally. Therefore, we proposed that Pfm-miR-9b-5p participated in shell formation by negatively regulating the expression of biomineralization gene PNU9 in Pinctada fucata martensii and this research provided some data to better understand the mechanisms of shell formation.
作者
李智鑫
熊新威
郑哲
黄荣莲
杜晓东
Li Zhixin;Xiong Xinwei;Zheng Zhe;Huang Ronglian;Du Xiaodong(Guangdong Provincial Engineering Laboratory for Mariculture Organism Breeding,Guangdong Science and Innovation Center for Pearl Culture,Pearl Breeding and Processing EngineeringTechnology Research Centre of Guangdong Province,Fishery College,Guangdong Ocean University,Zhanjiang,524088)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2022年第5期960-969,共10页
Genomics and Applied Biology
基金
广东省基础与应用基础研究基金项目(2019A1515011096、2020A1515010691)
国家自然科学基金项目(32002369)
广东省教育厅项目(2020ZDZX1045)共同资助
作者简介
通信作者:杜晓东,gdhddxd@hotmail.com
