Next-generation small RNA sequencing for microRNAs profiling in the honey bee Apis mellifera
Next-generation small RNA sequencing for microRNAs profiling in the honey bee Apis mellifera
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用于蜜蜂 Apis mellifera 中 microRNA 分析的新一代小 RNA 测序
DOI：
10.1111/j.1365-2583.2010.01039.x
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发表时间：
2010-12-01
影响因子：
2.6
通讯作者：
Hu, F.
中科院分区：
文献类型：
--
作者：
Chen, X.;Yu, X.;Hu, F.
MicroRNAs （miRNAs）通过转录后调控基因的表达，在各种生理和病理过程中起着关键的调节作用。蜜蜂（Apis mellifera）是高度社会性物种的关键模型，其复杂的社会行为可以从理论上解释为基因调控的变化，其中认为mirna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文库进行测序来鉴定蜜蜂的mirna库。共获得36 796 459个原始序列；其中5 491 100个短序列是mRNA和其他非编码rna （ncRNA）的片段，1 759 346个reads映射到已知的mirna。我们预测了267个新的蜜蜂mirna，代表380 182个短读，包括14 107 583个基因组定位序列中的8个其他昆虫mirna。我们利用茎环反转录PCR （RT-PCR）对其中50个进行了验证，其中35个获得了PCR产物。跨物种分析显示，81个新的mirna在其他昆虫中具有同源物，表明它们是真实的mirna，具有相似的功能。本研究结果为研究蜜蜂发育过程中的mirna调控网络和种姓分化等有趣现象提供了基础。
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.