Annotation of the zebrafish genome through an integrated transcriptomic and proteomic analysis

Zebrafish organ proteomics was carried out for proteogenomic analysis. High resolution mass spectrometry-based proteomic profiling of 11 adult organs (eye, brain, liver, spleen, intestine-pancreas, ovary, testes, muscle, heart and head) and two developmental stages (embryos 48 and 120 hours post-fertilization) of zebrafish (SAT line) was carried out. Protein extracts were reduced and alkylated (iodoacetamide). Off-loine fractionation was carried out by SDS-PAGE and basic RPLC. MS analysis was cariied on Agilent 6540 Q-TOF. Spleen and Testis samples were also analyzed on Thermo Orbitrap LTQ Velos.

本研究开展斑马鱼器官蛋白质组学研究，用于蛋白基因组学（proteogenomic）分析。采用高分辨质谱技术，对斑马鱼SAT品系的11种成体器官（眼、脑、肝、脾、肠-胰腺、卵巢、精巢、肌肉、心脏及头部）以及两个发育阶段（受精后48小时与120小时的胚胎）进行了蛋白质组全景分析。对蛋白质提取物进行还原与烷基化处理（采用碘乙酰胺）；通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）与碱性反相液相色谱（basic RPLC）完成离线分级分离。质谱分析采用安捷伦6540 Q-TOF质谱仪完成；脾脏与精巢样本还额外采用赛默飞Orbitrap LTQ Velos质谱仪进行了分析。