| 重组日本血吸虫14-3-3蛋白的纯化和鉴定 |
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| 引用本文： | 钟政荣,沈继龙,胡元生,李小月,罗庆礼. 重组日本血吸虫14-3-3蛋白的纯化和鉴定[J]. 中国生物制品学杂志, 2007, 20(6): 454-456 |
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| 作者姓名： | 钟政荣 沈继龙 胡元生 李小月 罗庆礼 |
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| 作者单位： | 蚌埠医学院第一附属医院检验科 (蚌埠233004)安徽医科大学病原生物学教研室安徽省基因研究重点实验室(合肥230032)(钟政荣)，安徽医科大学病原生物学教研室安徽省基因研究重点实验室 (合肥230032)(沈继龙，胡元生，李小月，罗庆礼) |
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| 基金项目： | 国家自然科学基金;安徽省自然科学基金;安徽省教育厅自然科学基金 |
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| 摘 要： | 目的观察两次切胶、两次电洗脱的方法纯化重组日本血吸虫14-3-3(Sj14-3-3)蛋白的效果。方法采用两次切胶、两次电洗脱的方法(与经典的切胶、电洗脱有所不同),纯化Sj14-3-3蛋白,并与柱层析纯化方法进行比较。结果采用两次切胶、两次电洗脱方法纯化的目的蛋白条带单一,浓度达3mg/ml,而柱层析法纯化的蛋白浓度仅为1mg/ml。Western blot证实纯化目的蛋白的特异性。结论两次切胶、两次电洗脱法可获得高纯度、高特异性的目的蛋白,不失为一种经济有效的蛋白纯化方法。
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| 关 键 词： | 血吸虫 14-3-3蛋白 纯化 鉴定 |
| 文章编号： | 1004-5503（2007）06-454-03 |
| 收稿时间： | 2006-10-25 |
| 修稿时间： | 2006-10-25 |
Purification and Identification of Recombinant Schistosoma japonicum 14-3-3 Protein |
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| ZHONG Zheng-rong , SHEN Ji-long, HU Yuan-sheng, et al. Purification and Identification of Recombinant Schistosoma japonicum 14-3-3 Protein[J]. Chinese Journal of Bilogicals, 2007, 20(6): 454-456 |
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| Authors: | ZHONG Zheng-rong SHEN Ji-long HU Yuan-sheng et al |
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| Affiliation: | Department of Laboratory, The First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, China |
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| Abstract: | Objective To purify and identify recombinant Schistosoma japonicum(Sj)14-3-3 protein.Methods Purify recombinant Sj 14-3-3 protein by 2 times of gel cutting and 2 times of electroelution.Test the purified protein for concentration by Coomassie brilliant blue staining,purity by SDS-PAGE and specificity by Western blot,and compared with those purified by column chromatography.Results The purified protein showed a single band with relative molecular mass of about 32 500 on SDS-PAGE profile,and reached a concentration of 3 mg/ml,which was significantly higher than that of protein purified by column chromatography(1 mg/ml).Western blot showed good specificity of the purified protein.Conclusion Highly purified and specific Sj 14-3-3 protein was obtained by developing an economic and effective purification procedure consisting of 2 times of gel cutting and 2 times of electroelution. |
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| Keywords: | Schistosoma japonicum 14-3-3 protein Purification Identification |
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