Identification of conserved microRNAs in Bombyx mori (silkworm) and regulation of fibroin L chain production by microRNAs in heterologous system
Identification of conserved microRNAs in Bombyx mori (silkworm) and regulation of fibroin L chain production by microRNAs in heterologous system
复制标题
家蚕中保守 microRNA 的鉴定以及异源系统中 microRNA 对丝心蛋白 L 链产生的调节。
DOI：
10.1016/j.ibmb.2008.09.008
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发表时间：
2008-12-01
影响因子：
3.8
通讯作者：
Jin, Yongfeng
中科院分区：
文献类型：
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作者：
Cao, Jun;Tong, Chuanzhou;Jin, Yongfeng
MicroRNAs(MiRNAs)是一类广泛的非编码小RNA，在基因表达调控中具有重要作用。在家蚕中鉴定miRNAs的研究还很少。在这里，通过计算同源搜索的方法鉴定了41个保守的miRNAs，其中一些被筛选出来并得到了实验证实。描述了位置特异性核苷酸分布、进化保守性和聚集性组织等特征。通过一般的miRNA靶标鉴定方法，我们还发现至少有4个miRNAs(miR-33/-190/-276/-7)靶向丝素蛋白L链转录本，这表明miRNAs可能在丝蛋白合成的调控中发挥重要作用。为了验证这一点，我们克隆了丝素蛋白L链基因的3‘端非编码区，并构建了几个表达载体。半反转录-聚合酶链式反应、组织化学和生物化学检测结果表明，miRNAs可以通过序列互补抑制报告基因(GUS)的表达。显然，农业渗透被证明是一种高效的方法来鉴定在异源系统中抑制其靶mRNA的miRNAs的功能。(C)2008爱思唯尔有限公司。保留所有权利。
MicroRNAs (miRNAs) are an extensive class of small non-coding RNAs with important roles in the regulation of gene expression. Little study has been performed on identifying miRNAs in Bombyx mori (silkworm). Here, 41 conserved miRNAs were identified by a computational homology search approach and some of them were selected and confirmed experimentally. Some characteristics such as position-specific nucleotide distribution, evolutionary conservation and clustered organization have been described. Through general method of miRNA target identification, we also find that at least 4 miRNAs (miR-33/-190/-276/-7) target fibroin L chain transcripts, suggesting that miRNAs might play an important role in the regulation of silk protein production. In order to validate this, 3'-UTR of fibroin L chain mRNA was cloned and several expression vectors were constructed. Assays of semi-RT-PCR, histochemical and biochemical assays after agroinfiltration indicated that miRNAs can repress reporter gene (GUS) expression with sequence complement. Clearly, agroinfiltration proved a highly efficient method to identify the function of miRNAs inhibiting their targets mRNA in heterologous system. (C) 2008 Elsevier Ltd. All rights reserved.