沙漠小球藻转人乳铁蛋白的电转优化

Optimization of Electro-Transformation Conditions for the Transformation HumanLactoferrinof Desert Chlorella

DOI：10.3969/j.issn.1673-1689.2018.09.004

中文关键词: 电转化 小球藻 人乳铁蛋白 正交实验 细胞致死率 荧光百分率 PCR阳性率

英文关键词: electroporation,Chlorella,human lactoferrin,orthogonal test,cell viability,fluorescence positive rate,PCR positive rate

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中文摘要:

对沙漠小球藻建立高效的遗传转化方法，对电击转化的条件和相关参数进行优化。作者以人乳铁蛋白基因作为电转条件优化的参考基因，采用单因素实验确定转染效率的电转化条件，然后进行电转化正交实验，得到最优的电转化组合。结果表明：最优的电转化组合是电压1 400 V、培养周期8 d、渗透剂浓度0.2 mol/L、渗透时间为0 h、质粒质量浓度为6 ug/mL、电击缓冲液浓度为0.4 mol/L，其细胞致死率为51.30%、荧光百分率为52.54%、PCR阳性平均百分率为42.30%。

英文摘要:

To establish an efficient genetic transformation protocol for DesertChlorella. It was optimized methods for electroporation conditions and related parameters. Human lactoferrin gene regard as reference gene of optimization of electro-transformation conditions. The various parameters of cell viability and transfection efficiency were determined with single factor experiments.Then，the optimal electroporation conditions were obtained through orthogonal test. Results shows the optimal composition of electroporation conditions：breakdown voltage 1 400 V，cell cycle 8day，osmosis buffer 0.2 mol/L，osmosis time 0 h，plasmid 6 g/mL，electroporation buffer 0.4 mol/L. Under the optimal composition of electroporation conditions，the detected results of cell viability is 51.30%，and fluorescence positive rate is 52.54%，and PCR positive rate is 42.30%.

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