海栖热袍菌电转化技术的研究

Electrotransformation of Thermotoga maritima MS8

[目的]对海栖热袍菌电转化技术进行研究。[方法]采用不同参数即不同的方型电脉冲参数、指数衰退型电脉冲参数和高电压电击参数对海栖热袍菌感受态细胞进行电转化,得到的转化子进行PCR验证。[结果]试验中发现以150 V的低压方形电脉冲,室温厌氧条件下电击25 ms、1次,有利于T.maritima MS8的转化,但这种以整合质粒进行的转化效率极低。[结论]为提高T.maritima MS8的基因转化效率奠定了基础。

[Objective]The aim was to study on the electrotransformation of Thermotoga maritima MS8.[Method] Square waves,exponential waves and high voltage shock were used for the electrotransformation of T.maritima MS8,and the obtained transformants were detected by PCR.[Result]A single square electric pulse could be applied to cell sample in 0.2 cm ET cuvettes by using a Bio-Rad Gene Pulser set at 150 V,25 ms at room temperature,but the transformation efficiency was very low.[Conclusion]This research may improve the transformation efficiency of T.maritima MS8.