猪源ETEC F18黏附素三种提取方法对照

Comparison of three different methods of extraction of ETEC F18 Fimbriae from Pigs

关键词:  F18黏附素  切割法  热洗脱法  联合热洗脱与切割法  SDS-PAGE

Key words:  F18 Fimbriae  homogenate technique  heat extraction  combined method of heat extraction and homogenate technique  SDS-PAGE

摘要:

[摘要] 为探索更为有效的黏附素提取技术，本试验以猪源ETEC F18+菌株为材料，通过三种方法（切割法、热洗脱法及联合热洗脱与切割法）各提取200mL菌悬液中的黏附素，应用紫外－可见分光光度计测定浓度并进行SDS-PAGE分析。结果显示，应用切割法、热洗脱法和联合热洗脱与切割法所得黏附素量分别为5.94mg、8.84mg和22.47mg，即应用联合热洗脱与切割法提取的黏附素量明显高于单纯应用热洗脱法和切割法；SDS-PAGE分析表明，联合热洗脱与切割法提取的黏附素蛋白纯度稍低于单纯应用热洗脱法和切割法。综合分析上述结果可知，联合热洗脱与切割法更适用于大量黏附素抗原的提取。

Abstract:

Abstract：To study the diversity of three different methods, homogenate technique, heat extraction and combined method of heat extraction and homogenate technique were applied to extract fimbriae of ETEC F18+ from pigs in this paper. Concentration and purity of the extractions were detected by ultraviolet-visible spectrophotometer analysis and SDS-PAGE. The results showed that the maximum quantity of fimbriae extracted from 200mL bacterial suspension by homogenate technique, heat extraction and combined method was 5.94, 8.84 and 22.47mg respectively, namely quantity of fimbriae extracted by combined method was the highest of the three. SDS-PAGE revealed that the purity of fimbriae extracted by combined method was slightly lower than that of either single method. Therefore the combined method was supposed to be the most suitable fimbriae extract method on a large scale, working efficiency and extraction quality together taken into account.