苏云金杆菌制剂毒素蛋白定量分析

本文研究了苏云金杆菌制剂中毒素蛋白的 SDS- PAGE-洗脱比色定量分析方法。B.t.样本用 Na OH溶液处理 ,然后加 L aemmli缓冲液处理 ,离心 ,取上清液进行电泳。用考马斯亮兰 (CBB) R- 2 50染色后 ,刮下130 KD-毒素蛋白区带 ,用吡啶洗脱蛋白吸附的 CBB R- 2 50 ,于 6 0 5nm测定溶液的吸光度 ,间接对毒素蛋白进行定量。本方法准确度高、精密度好。 RSD在 5%左右。添加回收率介于 94 .6 %～ 99.2 %。本方法与 SDS-PAGE- TL C-扫描法结果相比没有显著差异。 130 KD-毒素蛋白的量与生物测定得到的效价之间也具有较好的相关性。

QUANTITATIVE ANALYSIS OF TOXIN PROTEINS IN BACILLUS THURINGIENSIS PRODUCTS

The quantitative method, Sodium Dodecyl Sulphate Polyacryamide Gel Electrophoresis (SDS-PAGE)-Colorimetry, of toxin proteins in various B.t. commercial products was presented. B.t. samples were pretreated with aqueous NaOH at high pH, and treated with Laemmli sample buffer, then electrophoresis was carried out. After stained with Coomassie Brilliant Blue (CBB) R-250, the band of 130KD-toxin proteins was cut out, and the absorbed CBB R-250 was eluted with pyridine. The absorbance of the eluted dye was measured at 605nm. This method characterized by relatively high accuracy and precision. Relative standard deviation (RSD) was nearly 5%. The fortified recoveries varied from 94.6%～99.2%. There were no significant differences between the data from SDS-PAGE-Colorimetry and SDS-PAGE-TLC-Scanner. The relationship between the percentage of 130KD-toxin protein and the potency of insect bioassay was studied.