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重组人B淋巴细胞刺激因子原核表达条件的优化 被引量：2

The Expression Optimization of Recombinant Human B LympHocyte Stimulator (rhBLyS)in E.coli.
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摘要 　以重组人B淋巴细胞刺激因子(rhBLyS)蛋白表达宿主菌株M15(pQE30a/rhBLyS)作为研究对象,借助SDS PAGE分析方法,对于用IPTG诱导的重组目的产物的表达进行了优化研究,分析比较了pH值、裂解液种类、诱导时间、IPTG浓度、温度等参数对重组产物表达的影响.实验结果表明,降低温度和IPTG浓度有利于增加靶蛋白的可溶性,27℃和0 075mol/LIPTG为比较适合的表达条件. The expression optimization of recombinant human B lympHocyte stimulator (rhBLyS) gene in E.coli M15(DE3) using IPTG as the inducer with SDSPAGE method was studied.The influences of culture condition such as pH,lysis buffer,period of induction,IPTG concentration and temperature et al on expression of recombinant protein were compared and analyzed in detail.The results suggested that the declination of temperature and IPTG concentration contributed to the solubility of target protein and the highest expression level of total target protein achieved when induced for 4?h with 01?mol/L IPTG.The highest expression level of soluble rhBLyS obtained at 0075?mol/L IPTG and 27℃.
作者 杜秋丽 张双全 刘晓宇
机构地区 南京师范大学生命科学学院
出处 《南京师大学报（自然科学版）》 CAS CSCD 2003年第2期69-73,共5页 Journal of Nanjing Normal University(Natural Science Edition)
基金 南京师范大学科技创新基金(184070H80901).
关键词 重组人B淋巴细胞刺激因子 原核表达 rhBLyS 重组蛋白 肿瘤坏死因子 表达条件 免疫调控 rhBLyS,induce,IPTG
分类号 R394 [医药卫生—医学遗传学] Q786 [生物学—分子生物学]
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参考文献7

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同被引文献17

  • 1萨姆布鲁克J 弗里奇EF 曼尼阿蒂奇T 金冬雁 黎孟枫译.分子克隆实验指南[M] 第二版[M].北京:科学出版社,1992.880.
  • 2Kweon D H, Han N S, Park K M, et al. Overproduction of Phytolacca Insularis Protein in Batch and Fed-batch Culture of Recombinant Escherichia coli [J]. Process Biochem., 2001, 36(6): 537-542.
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  • 9奥斯伯F,金斯顿RE,赛德曼JG,等.精编分子生物学实验指南[M].第二版,北京:科学出版社,1998.332-333.
  • 10Gramajo HC,White J,Hutchinson CR,et al.Overproduction and localization of components of the polyketide synthase of Streptomyces glaucescens involved in the production of the antibiotic tetracenomycin C[J].J Bacteriol,1991,173 (20):6475-6483.

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南京师大学报（自然科学版）
2003年 第2期

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