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重组铜绿假单胞菌外毒素A工程菌的高密度发酵与表达
引用本文:董勇前,宣俊文,冯宜扬,丁浩,范锋锋,胡海涛,赵志强. 重组铜绿假单胞菌外毒素A工程菌的高密度发酵与表达[J]. 微生物学免疫学进展, 2011, 39(1): 24-27
作者姓名:董勇前  宣俊文  冯宜扬  丁浩  范锋锋  胡海涛  赵志强
作者单位:兰州生物制品研究所,兰州,730046
摘    要:高密度培养表达大肠杆菌生产重组铜绿假单胞菌外毒素A(rEPA)。用上海高机公司的30L自控发酵罐,采用分批补料培养技术,维持葡萄糖的浓度始终处于较低的水平,并分批补加氮源,同时溶氧控制在30%~40%,pH自动调控至7.0,培养至对数中期进行诱导。重组菌最终发酵液光密度(A600)未诱导时达到44,诱导时达到36,在上清液中表达的rEPA蛋白的含量占总蛋白的28.1%,在菌体中表达蛋白含量占总蛋白的4.7%。本实验为rE-PA的大规模生产奠定了基础。

关 键 词:铜绿假单胞菌外毒素A  高密度培养  表达

High cell density culture of recombinant E.coli to produce Pseudomonas aeruginosa exoprotein A
DONG Yong-qian,XUAN Jun-wen,FENG Yi-yang,et al.. High cell density culture of recombinant E.coli to produce Pseudomonas aeruginosa exoprotein A[J]. Progress In Microbiology and Immunology, 2011, 39(1): 24-27
Authors:DONG Yong-qian  XUAN Jun-wen  FENG Yi-yang  et al.
Affiliation:DONG Yong-qian,XUAN Jun-wen,FENG Yi-yang,et al.(Lanzhou Institute of Biological Products,Lznzhou 730046,China)
Abstract:To culture recombinant E.coli in high cell density to produce Pseudomonas aeruginosa exoprotein A(rEPA).30L autocontrol fermenter made by Shanghai GaoJi Corperation and fed-batch culture technology were used in this experiment,adding nitrogen nutrition in batches,keeping glucose concentration at low level,dissolved oxygen at 30% to 40%,pH at about 7.00 throughout the culture and induced at middle stage of logarithm growth stage.When not induced,the final cell density is 44,when induced,the final cell densit...
Keywords:Pseudomonas aeruginosa exoprotein A  High cell density culture  Expression  
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