水产学报2017，Vol.41Issue(9)：1345-1354,10.DOI:10.11964/jfc.20160910555
青虾冷休克蛋白Y-box编码基因的cDNA全长克隆与表达分析
Molecular cloning and expression analysis of cold shock protein Y-box gene from oriental river pawn (Macrobrachium nipponense)
摘要
Abstract
The Y-box protein family is highly conserved nucleic acid binding proteins which are conserved from bacteria to human.In this study,the eDNA encoding cold shock protein Y-box from oriental river pawn Macrobrachium nipponense (MnY-box) was cloned by using EST analysis and rapid amplification of eDNA ends (RACE) approach.The full length eDNA of MnY-box was 1501 bp,consisting of a 5'untranslated region of 84 bp,a 3'untranslated region of 541 bp,and an open reading frame of 876 bp.The deduced protein had 291 amino acid residues with molecular mass of 31.48 kDa.Based on the protein similarity comparison,the Y-box protein family was very conservative in CSD domain.Phylogenetic tree analysis stated that oriental river prawn has the closest relationship with Daphnia pulex.Quantitative real-time RT-PCR analysis showed that the MnY-box gene was expressed in haemocytes,hepatopancreas,muscles,gill,brain,testis,ovary and intestines with the highest level of expression in the hepatopancreas.After environmental low temperature and recovery challenge,the relative expression level of MnY-box in hepatopancreas was significantly higher compared to the control group at 3 h to 12 h post-low temperature,followed by return to control levels at recovery 12 h.What's more,we constructed prokaryotic expression system and obtained the recombinant proteins ofM.nipponense Y-box,which may provide information for further functional study.关键词
青虾/冷休克蛋白/低温/低氧/原核表达Key words
Macrobrachium nipponense/cold shock protein/low temperature/hypoxia/prokaryotic expression分类
生物科学引用本文复制引用
孙盛明,傅洪拓,戈贤平,朱健,乔慧,金舒博,张文宜..青虾冷休克蛋白Y-box编码基因的cDNA全长克隆与表达分析[J].水产学报,2017,41(9):1345-1354,10.基金项目
国家自然科学基金(31402280,31672633) （31402280,31672633）
国家“十二五”科技支撑计划(2012BAD25B07)National Natural Science Foundation of China (31402280,31672633) （2012BAD25B07）
National "Twelfth Five-Year" Plan for Science & Technology Support (2012BAD25B07) （2012BAD25B07）
