鳞头犬牙南极鱼atp6v0c基因在HeLa细胞中抗寒功能的研究  

Cloning of atp6v0c gene from Antarctic notothenioid fish Dissostichus mawsoni and identification of its cold tolerance in He La cell

作　　者：刘明丽[1] 王金凤[1] 张东[1] 陈良标[1] 

机构地区：[1]上海海洋大学水产与生命学院省部共建水产种质资源发掘与利用教育部重点实验室,上海201306

出　　处：《大连海洋大学学报》2016年第1期7-12,共6页Journal of Dalian Ocean University

基　　金：国家"973"重点基础研究发展计划项目(2010CB126304);国家基金委重点项目(31130049);上海市一流学科水产基础生物学项目

摘　　要：为探求南极鱼ATP酶类在低温适应下的作用,从鳞头犬牙南极鱼Dissostichus mawsoni的c DNA文库中克隆atp6v0c基因(dmatp6v0c),并转染到He La细胞中,通过流式细胞仪检测细胞在低温胁迫下(10℃)的死亡率,获得了dmatp6v0c基因的全部编码序列,其长度为462 bp,并将其插入到真核表达载体pc DNA3.1(-)上用于体外表达。结果表明,与对照组相比,在低温胁迫下,过表达dmatp6v0c基因的He La细胞死亡率从(19.23±1.87)%显著降低到(8.13±0.04)%(P<0.05)。研究表明,在低温胁迫下dmatp6v0c基因过表达能显著降低He La细胞的死亡率,试验结果可为dmatp6v0c基因作为鱼类耐寒育种的候选基因提供理论依据。The atpev0c gene of Antaratic notothenioid fish Dissostichus mawsoni was cloned from Antarctic notothe- nioid fish eDNA library, and then transfected the dmatp6vOc gene obtained into the HeLa cell to detect cell viability under cold stress （ 10 ℃ ） by flow cytometry to investigate the ATPase responsive for the cold adaptation in Antarc- tic notothenioid fish. The complete coding sequence with 462 bp of dmatp6vOc gene was cloned and sequenced, and the dmatp6vOc coding sequence was inserted into the pcDNA3.1 （-） vector by XhoI and BamHI restriction sites for gene expression in HeLa cell. The results showed that over-expression of dmatp6vOc gene in HeLa cell was found to reduce the cell mortality from 19.23% ±1.87% to 8.13% ±0.04% （P〈0.05） under cold stress com- pared with the control group. The findings indicated that the expression level of dmatp6vOc was heavily involved in cell viability under cold stress and that dmatp6vOc could be considered as a candidate for breeding cold resistant fish.

关 键 词：鳞头犬牙南极鱼 低温胁迫 dmatp6v0c基因 HELA细胞 细胞死亡率 

分 类 号：Q291[生物学—细胞生物学]