| 包涵体蛋白纯化方法的探讨 |
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| 引用本文： | 陈颖,沈敏,刘宝林.包涵体蛋白纯化方法的探讨[J].放射免疫学杂志,2011,24(1):62-64. |
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| 作者姓名： | 陈颖 沈敏 刘宝林 |
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| 作者单位： | 陈颖,Chen Ying(上海理工大学医疗器械与食品学院;上海杨浦区安图医院检验科,200092);沈敏,Shen Min(上海杨浦区安图医院检验科,200092);刘宝林,Liu Bao-lin(上海理工大学医疗器械与食品学院) |
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| 摘 要： | 目的:探讨包涵体蛋白的表达条件及纯化方法.方法:将PQE30-Cystatin转化大肠杆菌M15,通过IPTG诱导蛋白表达.为了能够获得可溶的且有活性的Cystatin C(Cys C),本文摸索了Cys C包涵体的复性条件及蛋白表达条件.结果:改变缓冲体系pH值及离子强度等多种方法均不能减少蛋白在复性过程中的沉淀;同...
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| 关 键 词： | 半胱氨酸蛋白酶抑制剂C 包涵体 可溶性表达 |
Study on Method of Purification of Inclusion Body Protein |
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| Chen Ying,Shen Min,Liu Bao-lin.Study on Method of Purification of Inclusion Body Protein[J].Journal of Radioimmanology,2011,24(1):62-64. |
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| Authors: | Chen Ying Shen Min Liu Bao-lin |
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| Institution: | Chen Ying~(1,2),Shen Min~2,Liu Bao-lin~1 1 Shanghai Ligong University School of Medical Equiment and Food,Shanghai(200093),China 2 Department of Clinical Laboratory,Autu Hospital,China |
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| Abstract: | Objective To study the optimal conditions for expression of inclusion body protein and method of purification of the protein.Methods PQE30-Cystatin C was transformed into E-Coli M15 and protein expression was induced with IPTG.Options for better renaturation of the obtained inclusion body protein were studied.Results We tried various ways including changes of buffer system pH and ionic strength,as well as OD 600 for protein expression,culture media pH,expression time……but failed to obtain soluble expression protein.However,we lowered the concentration of IPTG solution from 1mmol/L to 0.005mmol/L and finally obtained a slight amount of expressed soluble fusion Cystatin C protein.Conclusion Presently,the amount of soluble protein obtained with low concentration of IPTG is too little for practical use and further study is needed. |
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| Keywords: | Cystatin C(Cys C) inclusion body soluble expression |
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