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cDNA文库中克隆atp6v0c基因(dmatp6v0c),并转染到HeLa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pcDNA3.1(-)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HeLa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HeLa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 atp6 v0 c gene of Antaratic notothenioid fish Dissostichus mawsoni was cloned from Antarctic notothe-nioid fish cDNA library, and then transfected the dmatp6v0c 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 dmatp6v0c gene was cloned and sequenced, and the dmatp6v0c 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 dmatp6v0c 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 dmatp6v0c was heavily involved in cell viability under cold stress and that dmatp6 v0 c could be considered as a candidate for breeding cold resistant fish.
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