草鱼核因子E2相关因子2基因克隆分析及其 对呼吸爆发的调控作用
Cloning and analysis of nuclear factor E2-related factor 2 (Nrf2) and its function in the regulation of respiratory burst in grass carp (Ctenopharyngodon idella)
摘要
Abstract
Nuclear factor E2-related factor 2 (Nrf2) plays a key role in the regulation of respiratory burst which can produce amount of reactive oxygen species (ROS). In the present report, we have cloned and analyzed the Nrf2 gene from grass carp (Ctenopharyngodon idella), generated its polyclonal antibody, and characterized its role in regulation of ROS. The obtained cDNA of Nrf2 gene was 1994 bp, containing an open reading frame with 1782 bp encoding 593 amino acids. The amino acid sequences of Nrf2 from grass carp were highly identical with those from common carp (Cyprinus carpio) with 87％ similarity. Nrf2 from grass carp contained six typical Neh (Nrf2-Epoxy chloropropane (ECH) homology) domains, indicating it was highly conserved in evolution. The qRT-PCR and Western blot analysis showed that Nrf2 was expressed in all eight detected tissues. The total antioxidant capa-city of CIKs were up-regulated but ROS generated from CIK cells were down-regulated and the mRNAs of Nrf2 and its downstream gene (GST and HO-1) were all up-regulated when CIK was exposed to the ROS inducer, tert-Butylhydroquinone (tBHQ). Similarly, the expression of Nrf2 protein was also increased and it was translocated in the nucleus of CIK exposed to tBHQ. In conclusion, evolutionary conserved Nrf2 was ubiquitously expressed in the tissues of grass carp and it is involved in down-regulating the generation of ROS in CIK cells via up-regulating the expression of itself and its downstream antioxidant genes, so as to regulate respiratory burst.关键词
草鱼/核因子E2相关因子2(Nrf2)/基因克隆/多克隆抗体制备/呼吸爆发Key words
Ctenopharyngodon idella/NF-E2-related factor 2/gene cloning/polyclonal antibody/ROS分类
生物科学引用本文复制引用
冯燕,秦真东,代云佳,张玉蕾,刘小玲,周洋,兰江风,赵丽娟,林蠡..草鱼核因子E2相关因子2基因克隆分析及其 对呼吸爆发的调控作用[J].水产学报,2018,42(2):161-177,17.基金项目
国家自然科学基金(31502199,31572657,31372563) （31502199,31572657,31372563）
广东省海洋与渔业局基金(A201512C003,2015-115) （A201512C003,2015-115）
湖北省科技支撑计划(2015BBA228, YSF2015000255) （2015BBA228, YSF2015000255）
武汉市科技局资助课题(2016020101010089) （2016020101010089）
中央高校基本科研业务费专项(2014PY035) （2014PY035）
National Natural Science Foundation of China (31602190, 31572657, 31372563) （31602190, 31572657, 31372563）
Funds from the Administration of Ocean and Fisheries of Guangdong Province, China (A201512C003, 2015-115) （A201512C003, 2015-115）
Special Fund for Science and Technology from Hubei Province,China (2015BBA228, YSF2015000255) （2015BBA228, YSF2015000255）
Fund from Wuhan Science and Technology Bureau (2016020101010089) （2016020101010089）
Fundamental Research Funds for the Central Uni-versities (2014PY035) （2014PY035）
