College of Animal Sciences, Zhejiang University, Hangzhou, China.
Insect Mol Biol. 2010 Dec;19(6):799-805. doi: 10.1111/j.1365-2583.2010.01039.x. Epub 2010 Aug 30.
MicroRNAs (miRNAs) are key regulators in various physiological and pathological processes via post-transcriptional regulation of gene expression. The honey bee (Apis mellifera) is a key model for highly social species, and its complex social behaviour can be interpreted theoretically as changes in gene regulation, in which miRNAs are thought to be involved. We used the SOLiD sequencing system to identify the repertoire of miRNAs in the honey bee by sequencing a mixed small RNA library from different developmental stages. We obtained a total of 36,796,459 raw sequences; of which 5,491,100 short sequences were fragments of mRNA and other noncoding RNAs (ncRNA), and 1,759,346 reads mapped to the known miRNAs. We predicted 267 novel honey bee miRNAs representing 380,182 short reads, including eight miRNAs of other insects in 14,107,583 genome-mapped sequences. We verified 50 of them using stem-loop reverse-transcription PCR (RT-PCR), in which 35 yielded PCR products. Cross-species analyses showed 81 novel miRNAs with homologues in other insects, suggesting that they were authentic miRNAs and have similar functions. The results of this study provide a basis for studies of the miRNA-modulating networks in development and some intriguing phenomena such as caste differentiation in A. mellifera.
微小 RNA（miRNA）通过基因表达的转录后调控在各种生理和病理过程中起着关键的调节作用。蜜蜂（Apis mellifera）是高度社会化物种的重要模型，其复杂的社会行为可以从理论上解释为基因调控的变化，而 miRNA 被认为参与其中。我们使用 SOLiD 测序系统通过对来自不同发育阶段的混合小 RNA 文库进行测序，鉴定了蜜蜂的 miRNA 谱。我们总共获得了 36796459 条原始序列；其中 5491100 条短序列是 mRNA 和其他非编码 RNA（ncRNA）的片段，1759346 条读序列与已知的 miRNA 匹配。我们预测了 267 个新的蜜蜂 miRNA，代表 380182 个短读序列，包括在 14107583 条基因组映射序列中 14 种其他昆虫的 8 种 miRNA。我们使用茎环反转录 PCR（RT-PCR）验证了其中的 50 个，其中 35 个产生了 PCR 产物。种间分析显示，81 个新的 miRNA 在其他昆虫中有同源物，表明它们是真正的 miRNA，并具有相似的功能。这项研究的结果为 miRNA 调节网络在发育和一些有趣现象（如 A.mellifera 中的等级分化）中的研究提供了基础。
