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.
出处
《大连海洋大学学报》
CAS
CSCD
北大核心
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