重组大肠杆菌不耐热肠毒素B亚单位的纯化及生物学活性鉴定  

Purification and evaluation of the biological activity of recombinant E. coli heat-labile enterotoxin B

作　　者：田文标[1] 邹全明[1] 吴超[1] 张卫军[1] 杨珺[1] 冉向阳[1] 王缚鲲[1] 

机构地区：[1]第三军医大学医学检验系临床微生物学及免疫学教研室,重庆400038

出　　处：《第三军医大学学报》2003年第2期131-134,共4页Journal of Third Military Medical University

基　　金：国家"九五"科技攻关重点项目 ( 96 90 10 154 )

摘　　要：目的　纯化rLTB并对其生物学功能进行初步研究。方法　重组基因工程菌发酵后 ,以包涵体形式表达的rLTB经过洗涤、变性溶解后 ,分别采用QSepharoseTM HighPerformance阴离子交换层析和PhenylFF(highsub)疏水层析对其进行纯化并透析复性 ,然后采用SDS PAGE和HPLC检测纯度 ,并选用ELISA、动物实验和Western blotting实验对纯化蛋白的生物学活性进行鉴定。结果　得到纯度达 97 85 %的rLTB ,复性的rLTB经检测具有良好的生物学活性。结论　得到纯度较高、生物学活性较好的rLTB 。Objective To purify and evaluate the activity of recombinant E. coli heat labile enterotoxin B.Methods The inclusion body of the LTB expressed in E. coli was washed, denatured and renatured. The two step chromatographic procedure, Q Sepharose TM High Performance and Phenyl FF(high sub), was used for the purification. The purity of the rLTB was detected by SDS PAGE and HPLC, and the specific biological activity of the purified rLTB was detected by enzyme linked immunosorbent assay, animal test and Western blotting. Results The purity of rLTB was up to 97.85% and the renatured rLTB had a high specific biological activity.Conclusion The prepared rLTB with high purity and biological activity can be applied for further studies.

关 键 词：大肠杆菌 不耐热肠毒素 B亚单位 纯化 生物学活性 

分 类 号：R378.21[医药卫生—病原生物学]