Qi Jie, Liu Xudong, Liu Jinxiang, Yu Haiyang, Wang Wenji, Wang Zhigang, Zhang Quanqi
Key Laboratory of Marine Genetics and Breeding, Ocean University of China, 266003 Qingdao, Shandong, China.
Key Laboratory of Marine Genetics and Breeding, Ocean University of China, 266003 Qingdao, Shandong, China.
Fish Shellfish Immunol. 2014 Aug;39(2):503-11. doi: 10.1016/j.fsi.2014.05.038. Epub 2014 Jun 9.
Ambient temperature is one of the major abiotic environmental factors determining the main parameters of fish vital activity. HSP70 plays an essential role in heat response. In this investigation, the promoter and structure of Paralichthys olivaceus hsp70 (Pohsp70) gene was cloned and predicted. 2558 bp upstream regulatory region of Pohsp70 was annotated with four potential promoter elements and four putative binding sites of transcription factors heat shock elements (HSE, nGAAn) in the upstream of the transcription start site. In addition, one intron with 454 bp in the 5'-noncoding region was found. Quantitative Real Time PCR analysis indicated that the transcript level of Pohsp70 was raised markedly after 1 h by heat shocked. Furthermore, 25 SNPs were identified in Pohsp70 by resequencing, seven of which was associated with heat resistance. In addition, two of the seven SNPs, namely SNP14 and SNP16, were observed in strong linkage disequilibrium. The haplotype with association analysis showed TAGGAG haplotype was more represented in heat susceptible group while (DEL/T) GAATA haplotype was more frequent in heat resistant group. The heat resistant SNPs and haplotype could be candidate markers potentially serving for selective breeding programs of Japanese flounder aimed at improving anti-stress and production.
环境温度是决定鱼类生命活动主要参数的主要非生物环境因素之一。热休克蛋白70（HSP70）在热反应中起重要作用。在本研究中，对褐牙鲆hsp70（Pohsp70）基因的启动子和结构进行了克隆和预测。Pohsp70基因转录起始位点上游2558 bp的调控区域含有四个潜在的启动子元件和四个假定的转录因子热休克元件（HSE，nGAAn）结合位点。此外，在5′非编码区发现了一个454 bp的内含子。实时定量PCR分析表明，热休克1小时后，Pohsp70的转录水平显著升高。此外，通过重测序在Pohsp70中鉴定出25个单核苷酸多态性（SNP），其中7个与耐热性相关。此外，在这7个SNP中，SNP14和SNP16处于强连锁不平衡状态。单倍型关联分析表明，TAGGAG单倍型在热敏感组中更常见，而（DEL/T）GAATA单倍型在耐热组中更常见。耐热性SNP和单倍型可能作为候选标记，用于旨在提高抗应激能力和产量的牙鲆选择性育种计划。
