【作者】 黄琳玲；

【导师】 陈树德；

【作者基本信息】 华东师范大学 ， 生物物理， 2005， 硕士

【摘要】 前期的工作已发现,生长对数前期的红豆杉细胞在30min的脉冲电场(50Hz,约10V/m)作用下,细胞生长没有明显变化但紫杉烷的含量提高了近30%。本文通过蛋白质组学研究脉冲电场作用后48小时中国红豆杉细胞在蛋白表达上的变化。共进行了3次二维电泳重复实验,经过相关因子分析证实具有较高的重复性。图像分析比较对照组和经过脉冲电场处理的实验组的二维电泳图谱,发现有较明显的差异。从蛋白质分子水平上,说明脉冲电场的刺激对红豆杉细胞的蛋白质表达水平有明显的影响。用基质辅助激光解析-飞行时间质谱(MALDI-TOFMS)和液相色谱-电喷雾质谱(ESI MS)对其中部分发生变化的蛋白进行分析鉴定。鉴定结果表明:核糖核酸蛋白粒、醛糖差向异构酶和铁氧还蛋白-NADP(+)还原酶这三个蛋白可能是与脉冲电场诱导相关的蛋白。更多还原

【Abstract】 A significant increase in intracellular accumulation of taxuyunnanine c (Tc) was observed by exposing the suspension cultures of Taxus chinensis in early exponential growth phase to a 30-min pulsed electric field (PEF, 50Hz, ≈10V/m) in former study. Proteomic analysis was used to research the changes of the protein expression of suspension culture of Taxus chinensis 48 hours after the pulsed electric field elicitation. The two dimensional gel electrophoresis (2-DE) experiments was repeated for 3 times, and proved to be reliable according to the correlation coefficient analysis. Distinct changes were found from comparison of the protein patterns of control and PEF treated cells. Matrix-assisted laser desorption/ionization mass spectrometry and electrospray ionization mass spectrometry were used to analyse some changed proteins. The conclusions indicated that heterogeneous nuclear ribonucleoprotein(hnRNP), aldose 1-epimerase family protein and Ferredoxin—NADP(+) reductase, putative/adrenodoxin reductase were related to the induction of PEF.更多还原