| 重组幽门螺杆菌粘附素的纯化工艺 |
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| 引用本文： | 刘正祥,邹全明,洪愉,朱永红.重组幽门螺杆菌粘附素的纯化工艺[J].粉末涂料与涂装,2004,17(3):145-148. |
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| 作者姓名： | 刘正祥 邹全明 洪愉 朱永红 |
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| 作者单位： | 第三军医大学检验系临床微生物及免疫学教研室,重庆400008 |
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| 基金项目： | 国家“九五”重点科技攻关课题(96-901-01-54)，“八六三”生物与现代农业技术领域生物工程技术主题课题(2001AA21516). |
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| 摘 要： | 目的纯化大肠杆菌表达的重组幽门螺杆菌粘附素。方法将表达HpaA蛋白的工程菌经高压均质机破菌获得包涵体,经洗涤、变性、复性,Q Sepharose High Performance阴离子交换层析和亲和层析分离纯化。采用SDS-PAGE和HPLC检测纯度,用Western blot检测其抗原性。结果纯化后HpaA蛋白纯度高达95％以上,具有良好的抗原性。结论建立了从包涵体中获得高纯度HpaA纯化工艺,为进一步的研究打下了基础。
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| 关 键 词： | 幽门螺杆菌鞭毛粘附素 包涵体 层析 |
| 修稿时间： | 2003年6月30日 |
Purification of Recombinant Helicobacter pylori Adhesin Protein HpaA |
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| LIU Zheng-xiang,ZOU Quan-ming,Hong Yu,et al.Purification of Recombinant Helicobacter pylori Adhesin Protein HpaA[J].Chinese Journal of Biologicals,2004,17(3):145-148. |
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| Authors: | LIU Zheng-xiang ZOU Quan-ming Hong Yu |
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| Abstract: | Objective To establish an effective method for the purification of recombinant Helicobact-er pylori adhesin protein (rHpaA) expressed in E. coli. Methods The recombinant strain expressing HpaA protein was disrupted by Hyperbric homogeniser. The obtained inclusion bodies were washed, de-naturalized and re-naturalized, and the HpaA in them were purified by Q Sepharose High Performance negative ion exchange and affinity chromatography. The purity of HpaA was detected by SDS-PAGE and HPLC, and its anti-genicity by Western blot.Results The purified HpaA protein reached a purity of above 95% and showed good antigenicity. Conclusion An procedure for high purification of HpaA from inclusion body was developed. |
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| Keywords: | Helicobacter pylori adhesin Inclusion body Charomatography |
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