| 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 |
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| 引用本文： | 田文标,邹全明,吴超,张卫军,杨珺,冉向阳,王缚鲲. 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[J]. 第三军医大学学报, 2003, 25(2): 131-134 |
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| 作者姓名： | 田文标 邹全明 吴超 张卫军 杨珺 冉向阳 王缚鲲 |
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| 作者单位： | 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 |
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| 基金项目： | 国家“九五”科技攻关重点项目 ( 96 90 10 154 ) |
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| 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Q Sepharose^TM High Performance阴离子交换层析和Phenyl FF(high sub）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，可用于进一步的研究。
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| 关 键 词： | 大肠杆菌 不耐热肠毒素 B亚单位 纯化 生物学活性 |
| 文章编号： | 1000-5404(2003)02-0131-04 |
| 修稿时间： | 2002-03-05 |
Purification and evaluation of the biological activity of recombinant E. coli heat-labile enterotoxin B |
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| TIAN Wen biao,ZOU Quan ming,WU Chao,ZHANG Wei jun,YANG Jun,RAN Xiang yang,QANG Fu kun. Purification and evaluation of the biological activity of recombinant E. coli heat-labile enterotoxin B[J]. Acta Academiae Medicinae Militaris Tertiae, 2003, 25(2): 131-134 |
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| Authors: | TIAN Wen biao ZOU Quan ming WU Chao ZHANG Wei jun YANG Jun RAN Xiang yang QANG Fu kun |
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| Abstract: | 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. |
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| Keywords: | heat labile enterotoxin inclusion body protein purification |
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