【目的】通过分析梅花鹿早期妊娠不同时期的血液转录组数据,筛选参与梅花鹿早期妊娠的关键基因。【方法】分别采集人工授精后0、7、15和20 d的雌性梅花鹿血液样品,利用Illumina HiSeq TM 3000C测序平台进行转录组分析。对不同时期的血...【目的】通过分析梅花鹿早期妊娠不同时期的血液转录组数据,筛选参与梅花鹿早期妊娠的关键基因。【方法】分别采集人工授精后0、7、15和20 d的雌性梅花鹿血液样品,利用Illumina HiSeq TM 3000C测序平台进行转录组分析。对不同时期的血液样本进行主成分分析和组间表达相关性分析,鉴定梅花鹿早期妊娠的关键阶段,利用DESeq筛选各时期的差异表达基因(DEGs),并进行GO和KEGG功能分析以及蛋白互作网络构建。【结果】转录组测序结果显示,妊娠第15天时梅花鹿血液出现了转录高峰,比较组D7 vs D0、D15 vs D7、D20 vs D15分别筛选到181、996、265个DEGs。其中比较组D7 vs D0共筛选到ERAS、MET、KDR等7个关键基因。综合GO和KEGG功能富集结果显示,基因主要富集在细胞对刺激的响应、信号转导等生物过程,以及PI3K-Akt信号通路、MAPK信号通路等通路中;比较组D15 vs D7共筛选到HRAS、HIF 1A、MAPK 1、JAK 1等10个关键基因,主要富集在免疫系统进程、免疫反应、趋化因子信号通路、C型凝集素受体信号通路等信号通路中;比较组D20 vs D15共筛选到MAPK 1、MMP 2等4个关键基因,主要富集在雌激素信号通路和松弛素信号通路中。【结论】本研究分析了梅花鹿早期妊娠关键时间点血液转录组的动态表达,筛选到KDR、HRAS、MAPK 1、JAK 1等关键基因,为梅花鹿早期妊娠诊断标志物的挖掘提供参考。展开更多
Early detection of pregnancy is advantageous to breeders. However, tools that were used in the early pregnancy detection are expensive, laboratory-based and not efficient and applicable enough in the field to farmers....Early detection of pregnancy is advantageous to breeders. However, tools that were used in the early pregnancy detection are expensive, laboratory-based and not efficient and applicable enough in the field to farmers. Thus, there is a need to find a biomarker which not only can detect pregnancy but also could be applied to the livestock on the field. Proteomic approach was used in this study as to search for early pregnancy biomarker. When goat sera at day 14 of gestation were resolved by using two dimensional gel electrophoresis, five differentially expressed proteins were detected. Four of the proteins were identified as albumin precursors, immunoglobulin lambda light chain and zinc-alpha-2-glycoprotein. Pregnancy associated glycoprotein-1 was detected by using liquid chromatography mass spectrometry and the protein was validated by immunoblotting. These proteins have potential to be used as a biomarker for early pregnancy detection, however, more extensive analysis are needed to validate its possibility.展开更多
文摘【目的】通过分析梅花鹿早期妊娠不同时期的血液转录组数据,筛选参与梅花鹿早期妊娠的关键基因。【方法】分别采集人工授精后0、7、15和20 d的雌性梅花鹿血液样品,利用Illumina HiSeq TM 3000C测序平台进行转录组分析。对不同时期的血液样本进行主成分分析和组间表达相关性分析,鉴定梅花鹿早期妊娠的关键阶段,利用DESeq筛选各时期的差异表达基因(DEGs),并进行GO和KEGG功能分析以及蛋白互作网络构建。【结果】转录组测序结果显示,妊娠第15天时梅花鹿血液出现了转录高峰,比较组D7 vs D0、D15 vs D7、D20 vs D15分别筛选到181、996、265个DEGs。其中比较组D7 vs D0共筛选到ERAS、MET、KDR等7个关键基因。综合GO和KEGG功能富集结果显示,基因主要富集在细胞对刺激的响应、信号转导等生物过程,以及PI3K-Akt信号通路、MAPK信号通路等通路中;比较组D15 vs D7共筛选到HRAS、HIF 1A、MAPK 1、JAK 1等10个关键基因,主要富集在免疫系统进程、免疫反应、趋化因子信号通路、C型凝集素受体信号通路等信号通路中;比较组D20 vs D15共筛选到MAPK 1、MMP 2等4个关键基因,主要富集在雌激素信号通路和松弛素信号通路中。【结论】本研究分析了梅花鹿早期妊娠关键时间点血液转录组的动态表达,筛选到KDR、HRAS、MAPK 1、JAK 1等关键基因,为梅花鹿早期妊娠诊断标志物的挖掘提供参考。
文摘Early detection of pregnancy is advantageous to breeders. However, tools that were used in the early pregnancy detection are expensive, laboratory-based and not efficient and applicable enough in the field to farmers. Thus, there is a need to find a biomarker which not only can detect pregnancy but also could be applied to the livestock on the field. Proteomic approach was used in this study as to search for early pregnancy biomarker. When goat sera at day 14 of gestation were resolved by using two dimensional gel electrophoresis, five differentially expressed proteins were detected. Four of the proteins were identified as albumin precursors, immunoglobulin lambda light chain and zinc-alpha-2-glycoprotein. Pregnancy associated glycoprotein-1 was detected by using liquid chromatography mass spectrometry and the protein was validated by immunoblotting. These proteins have potential to be used as a biomarker for early pregnancy detection, however, more extensive analysis are needed to validate its possibility.
