He Hua, Chen Si, Liang Wei, Liu Xiaolin
College of Animal Science and Technology, Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, China.
College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
Anim Genet. 2017 Apr;48(2):131-140. doi: 10.1111/age.12508. Epub 2016 Sep 19.
To gain further insight into the molecular mechanism of bovine muscle development, we combined mass spectrometry characterization of proteins with Illumina deep sequencing of RNAs obtained from bovine longissimus muscle (LD) at prenatal and postnatal stages. For the proteomic study, each group of LD proteins was extracted and labeled using isobaric tags for relative and absolute quantitation (iTRAQ) method. Among the 1321 proteins identified from six samples, 390 proteins were differentially expressed in embryos at day 135 post-fertilization (Emb135d) vs. 30-month-old adult cattle (Emb135d vs. 30M) samples. Gene Ontology, Cluster of Orthologous Groups and Kyoto Encyclopedia of Genes and Genomes analyses were further conducted to better understand the different functions. Furthermore, we analyzed the relationship between transcript and protein regulation between samples by direct comparison of expression levels from transcriptomic and iTRAQ-based proteomics. Association results indicated that 1295 of 1321 proteins could be mapped to transcriptome sequencing data. This study provides the most comprehensive, targeted survey of bovine LD proteins to date and has shown the power of combining transcriptomic and proteomic approaches to provide molecular insights for understanding the developmental characteristics in bovine muscle, and even in other mammals.
为了更深入了解牛肌肉发育的分子机制,我们将蛋白质的质谱表征与从牛产前和产后阶段的背最长肌(LD)获得的RNA的Illumina深度测序相结合。对于蛋白质组学研究,每组LD蛋白质均采用相对和绝对定量等压标签(iTRAQ)方法进行提取和标记。在从六个样本中鉴定出的1321种蛋白质中,390种蛋白质在受精后135天的胚胎(Emb135d)与30月龄成年牛(Emb135d与30M)样本中差异表达。进一步进行了基因本体论、直系同源簇和京都基因与基因组百科全书分析,以更好地了解不同的功能。此外,我们通过直接比较转录组学和基于iTRAQ的蛋白质组学的表达水平,分析了样本之间转录本与蛋白质调控的关系。关联结果表明,1321种蛋白质中的1295种可以映射到转录组测序数据。这项研究提供了迄今为止对牛LD蛋白质最全面、有针对性的调查,并展示了结合转录组学和蛋白质组学方法为理解牛肌肉甚至其他哺乳动物的发育特征提供分子见解的能力。