小鼠后肢特异表达基因的筛选及验证
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(东华大学生物研究所,上海 201620)

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Q95-33

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Screening and verification of mouse hind limb-specific gene expression
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(Institute of Biology,Donghua University,Shanghai 201620,China)

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    摘要:

    目的 对比小鼠前后肢基因表达谱的变化,筛选小鼠后肢特异表达的基因。方法 通过NCBI GEO数据库查找小鼠前后肢基因表达芯片,使用Transcriptome Analysis Console 软件对原始数据(5 个前肢E10. 5 d 芯片及4 个后肢E10. 5 d 芯片)进行质控,利用差异倍数为2 倍以上或0. 5 倍以下及 P 值小于0. 05 进行数据过滤,绘制差异表达基因的火山图,并进行聚类分析,构建后肢特异表达基因的相互作用分子网络图。结果 小鼠胚胎E10. 5d 后肢相较于前肢的差异表达的基因共有275 个,其中上调的基因45 个,下调的基因230 个,构建了后肢特异表达的差异基因分子网络图,找到后肢特异表达核心基因,使用qPCR 检测部分后肢特异表达的基因,结果与芯片结果一致。结论 应用小鼠表达谱芯片成功筛选出后肢特异表达的基因,为哺乳动物肢体发育的研究提供基础。

    Abstract:

    Objective To compare the gene expression profiles of fore and hind limbs in mice and screen out genes specifically expressed in hind limbs. Methods Mouse fore and hind limb gene expression chip data were obtained from the NCBI GEO database. The raw data (five forelimb E10. 5 day chips and four hind limb E10. 5 day chips) were quality-controlled using the Transcriptome Analysis Console software. The data were filtered to identify differentially expressed genes (DEGs) using the criteria: difference > 2 times or < 0. 5 times and P-value. We drew volcano maps and performed cluster analysis with the DEGs. A gene interaction network map was constructed with the hind limb-specific genes. Results There were 275 DEGs in the hind limbs of mouse E10. 5 embryos, of which 45 genes were upregulated and 230 genes were downregulated. The hind limb-specific DEGs were mapped to the gene network and the core node genes of hind limbs were identified. qPCR was used to investigate some of the hind limb-specific genes, and the results were consistent with the chip results. Conclusions Mouse expression microarrays are successfully used to screen the genes specifically expressed in hind limbs, which provide a basis for the study of mammalian limb development.

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王茂春,段维旺,李凯,周宇荀,肖君华.小鼠后肢特异表达基因的筛选及验证[J].中国实验动物学报,2019,27(1):1~6.

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  • 收稿日期:2018-09-04
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  • 在线发布日期: 2019-03-06
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