基于转录组测序技术挖掘大豆蛋白质合成相关基因
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摘要
通过转录组测序技术挖掘蛋白质合成相关基因,为解析蛋白质合成机制奠定基础。以遗传背景相近但蛋白含量差异较大的大豆高蛋白品系冀HJ117(蛋白质含量52. 99%)及其回交亲本冀豆12(蛋白质含量46. 48%)为研究材料,利用转录组测序技术和生物信息学分析,以期挖掘大豆籽粒蛋白质合成相关基因。通过对转录组数据中冀HJ117和冀豆12差异表达基因的分析筛选,共得到336个差异表达基因,其中冀HJ117较冀豆12有195个上调表达基因,141个下调表达基因。通过GO功能富集分析,发现在分子功能类型中注释的差异基因主要与催化和连接等功能相关; KEGG显著富集分析发现差异表达基因主要富集在蛋白质内质网合成途径中,该途径共筛选到34个差异表达基因,其中33个基因在冀HJ117中表达量较高。从该途径中筛选出9个候选基因,采用荧光定量PCR方法检测其表达量,表达趋势与RNA-Seq检测结果基本一致,证实了RNA-Seq数据的可靠性。RNA-Seq测序结果获得了与蛋白质合成相关基因的基本信息,蛋白质内质网合成途径可能是冀HJ117与冀豆12蛋白质含量产生差异的重要通路。
By using transcriptome sequencing technology,the screening of protein synthesis related genes will lay a foundation for further exploring the mechanism of protein synthesis. Ji HJ117( Protein content of 52. 99%)and its backcross parent Jidou 12( Protein content of 46. 48%) with similar genetic backgrounds but different in protein content were used as research materials. Through RNA-sequencing of soybean seeds from Ji HJ117 and Jidou12 to gain insights into the genes potentially related to protein synthesis. A total of 336 differentially expressed genes( DGEs) were screened from Ji HJ117 and Jidou 12,of which 195 were up-regulated and 141 down-regulated in Ji HJ117. Through the Gene Ontology analysis,DGEs annotated in molecular function types were mainly related to catalysis and ligation. Significant enrichment analysis of KEGG revealed that the DGEs were mainly enriched in protein processing in endoplasmic reticulum. A total of 34 DGEs were screened in this pathway,of which 33 genes were highly expressed in Ji HJ117. Nine candidate genes were selected from this pathway and their expression levels were detected by Real-time fluorescence quantitative PCR. The expression trend was basically consistent with that of RNA-Seq,confirming the reliability of RNA-Seq datas. RNA-Seq analysis obtained basic information about protein synthesis related genes in soybean. The pathway of protein processing in endoplasmic reticulum might be an important pathway that contribute to the difference of protein content in Ji HJ117 and Jidou 12. The accomplishment of these results constitute the first step toward understanding the regulation mechanisms of seed protein synthesis and provide valuable resource for further research.
By using transcriptome sequencing technology,the screening of protein synthesis related genes will lay a foundation for further exploring the mechanism of protein synthesis. Ji HJ117( Protein content of 52. 99%)and its backcross parent Jidou 12( Protein content of 46. 48%) with similar genetic backgrounds but different in protein content were used as research materials. Through RNA-sequencing of soybean seeds from Ji HJ117 and Jidou12 to gain insights into the genes potentially related to protein synthesis. A total of 336 differentially expressed genes( DGEs) were screened from Ji HJ117 and Jidou 12,of which 195 were up-regulated and 141 down-regulated in Ji HJ117. Through the Gene Ontology analysis,DGEs annotated in molecular function types were mainly related to catalysis and ligation. Significant enrichment analysis of KEGG revealed that the DGEs were mainly enriched in protein processing in endoplasmic reticulum. A total of 34 DGEs were screened in this pathway,of which 33 genes were highly expressed in Ji HJ117. Nine candidate genes were selected from this pathway and their expression levels were detected by Real-time fluorescence quantitative PCR. The expression trend was basically consistent with that of RNA-Seq,confirming the reliability of RNA-Seq datas. RNA-Seq analysis obtained basic information about protein synthesis related genes in soybean. The pathway of protein processing in endoplasmic reticulum might be an important pathway that contribute to the difference of protein content in Ji HJ117 and Jidou 12. The accomplishment of these results constitute the first step toward understanding the regulation mechanisms of seed protein synthesis and provide valuable resource for further research.
引文
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