人孕酮受体激素结合区片段在大肠杆菌中的表达条件

Expressing of LBD Segment of hPR in E·colic

本试验研究了人孕酮受体激素结合区片段在大肠杆菌中的表达条件 ,结果发现 ,随着诱导时间的延长 ,表达量增加 ,4h时 ,表达量最大 ,诱导时间进一步延长 ,表达量并未增加 ;当诱导剂浓度增加时 ,表达量增加 ,浓度为 1mmol/ L时 ,表达量最大 ,浓度进一步增加 ,表达量无明显变化 ;培养液中青霉素浓度在 10 0～ 180 μg/ m L 时表达量最大 ,进一步增加青霉素浓度 ,表达量开始下降。当浓度为 30 0μg/ m L时 ,仅见一条非常模糊的带。用 Western印迹检测时发现 ,在与 45k Da平行的位置 ,最适反应条件下的产物与不加 IPTG时相比 。

In order to acquire much more expressing product,We studed effects of inducing time,inducer concentration,concentration of ampicillin in the culture.With the addition of inducing time,the product began to increase.By the end of 4h,the product was the most.With the addition of inducer concentration,the product began to increase.In the concentration of 1 mmol/mL,the product was the most.From 100～180 μg/mL of ampicillin concentration in the culture,the product was the most and had no difference.Using western blot to deterct the production,we found there was a band in the postion of molecular weight 45 kDa.