黄河鲤肌间骨转录组分析及斑马鱼bmp1基因敲除模型构建

摘要：

肌间骨(IBs)由肌膈间的肌腱骨化而来 ,仅存在于低等硬骨鱼类中 ,研究选取黄河鲤(Cyprinuscarpio haematopterus)肌间骨发育前后两个关键阶段进行转录组测序 ,并对差异表达基因进行功能富集分析 . 结果表明 :测  序共获得高质量 有 效 序 列 120 356 780条 ,共 筛 选 出 差 异 表 达 基 因 3 454个(上 调 1 283个 , 下 调 2 171个) . GO 和  KEGG分析显示差异基因主要富集于胞外区域 、跨 膜 运 输 、氧 化 还 原 过 程 、催 化 活 性 、蛋 白 结 合 等 功 能 ,参 与 次 生 代  谢物的生物合成 、PI3K-Akt信号通路 、AMPK信号通路 、糖酵解等 . 经 qRT-PCR验证差异基因表达谱与 RNA-seq结  果基本一致后 ,为进一步了解其生物功能 ,选取两阶段中显著上调基因 bmp1,利用 CRISPR/Cas9建立 bmp1突变的  嵌合体斑马鱼(Daniorerio) . 测序验证 bmp1突变品系已 成 功 构 建 , 表 型 观 察 结 果 发 现 突 变 型 明 显 发 育 迟 缓 且 尾 部  出现脊椎畸形 .

Intermuscular bones(IBs)  arise from the ossification of the tendons between the muscle diaphragm and are found only in lowerbony fishes. In this study, transcriptome sequencing was performed in Cyprinuscarpio haematopterus at two key stagesofintermuscularbonedevelopment. Transcriptomedatawereanalyzed to identifydifferentially expressed genes,  and functionalenrichmentanalysiswasconducted forthem. Theresultsshowed thata totalof120356780highquality readse- quences were obtained, and 3 454 differentially expressed genes(1 283 up-regulated and 2 171 down-regulated) were screened.  ByGO and KEGG analysis, differentialgenes were mainly enriched in extracellular regions, transmembrane transport, RED- OX process, catalytic activity, protein binding and other functions, and involved in biosynthesis of secondary metabolites,  PI3K-Aktsignaling pathway, AMPK signaling pathway, glycolysis and so on.  After qRT-PCR verified that the differential gene expression profile was basically consistentwith the results ofRNA-seq, in orderto furtherunderstand its biologicalfunc- tion, the significantlyup-regulated genebmp1 wasselected in two stages, and CRISPR/Cas9 wasused to establish chimericze- brafish withbmp1 mutation. Sequencing confirmed thatthe mutantstrain ofbmp1had been successfully constructed, and phe- notypic observation showed thatthe mutanthad obvious developmentalretardation and vertebraldeformities in the tail.

作者：

董传举 , 贾颖颖 , 申亚伟,李学军

Dong Chuanjua, Jia Yingying, Shen Yaweia , LiXuejuna

机构地区：

betway官方app 水产学院;三农法律问题研究中心 

引用本文：

董传举,贾颖颖,申亚伟等.黄河鲤肌间骨转录组分析及斑马鱼bmp1基因敲除模型构建[J].betway官方app 学报(自然科学学版) ,2024,52(4)：71-79. 

Dong Chuanju,Jia Yingying,Shen Yawei,et al.Transcriptome analysis of intermuscularbonein Yellow RiverCarp and construction ofbmp1geneknockoutmodelin Zebrafish[J] .Journal of Henan Normal University(Natural Science Edition) , 2024, 52(4) : 71-79. DOI: 10. 16366/j. cnki. 1000-2367.  2023. 01. 11. 0002.

基金：

国家自然科学基金;河南省科技攻关项目 

关键词：

肌间骨;黄河鲤 ;转录组 ;bmp1;基因敲除

intermuscularbone; Yellow River carp; transcriptome; bmp1; gene knockout

分类号：

S917. 4  

黄河鲤肌间骨转录组分析及斑马鱼bmp1基因敲除模型构建.pdf