细胞饼瞬时表达体系的优化

Efficiency Optimization for the Cell Pack Transient Expression System

细胞饼瞬时转化技术具有重要的应用前景,然而其转化效率的优化研究尚未见报道。通过农杆菌介导法,系统地比较不同农杆菌浓度,抗氧化剂以及表面活性剂的添加对细胞饼瞬时转化效率的影响。研究结果表明:1)侵染用农杆菌的浓度在较高水平(OD=1.0)时更有利于转化效率的提升;2)两种抗氧化剂中,辛硫酸效果较半胱氨酸为好。但两者均表现为低浓度对转化效率具有促进作用,而高浓度则表现为抑制作用;3)测试的两种表面活性剂,Triton X-100以及Silwet L-77对转化效率均具有显著提升作用,而Silwet L-77的作用则更为突出;4)在优化条件下,人白介素10的含量较对照高出3.66倍。本研究的结果可进一步提升细胞饼瞬时转化技术的应用前景,为后续这一技术的应用奠定了一定的理论基础。

The transient expression of cell pack has important application prospects, but the optimization of its transformation efficiency has not been reported yet. In this study, the effects of different concentrations of Agro-bacterium tumefaciens, antioxidant and surfactant on transient expression efficiency of cell pack were systematically compared by A. tumefaciens-mediated transformation. The results showed that: 1. The concentration of A. tumefaciens for infection was more conducive to the improvement of transformation efficiency at a high level (OD= 1.0);2. Lipoic acid showed better performance than that of cysteine. Low concentration of antioxidant promoted the transformation efficiency, while high concentration showed inhibition effects;3. Both of the tested surfactants, Triton X-100 and Silwet L-77, had significant improvement on the transformation efficiency. While, the effect of Silwet L-77 was more prominent. 4. Under the optimized conditions, the content of human interleukin-10 was 3.66 times higher than that of the control. The results of this study can further enhance the application prospects of transient cell cake transformation technology, and lay a theoretical foundation for the follow-up application of this technology.