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An established kidney cell line from humpback grouper (Cromileptes altivelis) and its susceptibility to bacteria and heavy metals

机译:来自座头石斑鱼(Cromileptes altivelis)的已建立肾细胞系及其对细菌和重金属的敏感性

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摘要

Humpback grouper (Cromileptes altivelis), one kind of commercial fish with considerable economic value, has been recognized as a promising candidate for mariculture. In the wake of the development of aquaculture industry, the breeding density of C. altivelis has increased gradually, which gave rise to the occurrence of various pathogenic diseases. In our research, we established a new kidney cell line (designated as CAK) from humpback grouper and evaluated its susceptibility to bacteria and heavy metals. The results of our study showed that the optimal growth temperature was 26 degrees C, and optimal medium was L-15 supplemented with 20 fetal bovine serum (FBS). The sequencing of 18S rRNA gene indicated that CAK cell line was derived from C. altivelis. Chromosome analysis showed that the number of chromosome in CAK was 48. After being transfected of pEGFP-N3 plasmid, high transfection efficiency of CAK was observed, suggesting the potential to be used for the study of foreign functional genes. Moreover, the bacterial susceptibility results revealed that CAK cells were sensitive to Vibrio harveyi and Edwardsiella tarda, especially V. harveyi. Meanwhile, three heavy metals (Hg, Cu, and Cd) had toxic effects on the CAK cells with a dose-dependent manner. To sum up, the CAK cell line might be an ideal tool in vitro for analyzing the function of exogenous genes, bacterial susceptibility, and toxicity assay of heavy metals.
机译:座头石斑鱼(Cromileptes altivelis)是一种具有相当经济价值的商业鱼类,已被公认为海水养殖的有前途的候选者。随着水产养殖业的发展,白芷的养殖密度逐渐增加,引发了各种致病病害的发生。在我们的研究中,我们从座头石斑鱼中建立了一种新的肾细胞系(命名为CAK),并评估了其对细菌和重金属的敏感性。研究结果表明,最佳生长温度为26°C,最佳培养基为L-15,添加20%胎牛血清(FBS)。18S rRNA基因测序结果显示,CAK细胞系来源于C. altivelis。染色体分析显示,CAK中的染色体数量为48条。转染pEGFP-N3质粒后,观察到CAK的高转染效率,表明CAK具有用于研究外源功能基因的潜力。此外,细菌敏感性结果显示,CAK细胞对哈维弧菌和迟发性爱德华氏菌敏感,尤其是哈维弧菌。同时,3种重金属(Hg、Cu和Cd)对CAK细胞具有剂量依赖性毒性作用。综上所述,CAK细胞系可能是体外分析外源基因功能、细菌敏感性和重金属毒性测定的理想工具。

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