Stationary phase protein overproduction is a fundamental capability of Escherichia coli.
Although Escherichia coli is well studied and various recombinant E. coli protein expression systems have been developed, people usually consider the rapid growing (log phase) culture of E. coli as optimum for production of proteins. However, here we demonstrate that at stationary phase three E. coli systems, BL21 (DE3)(pET), DH5alpha (pGEX) induced with lactose, and TG1 (pBV220) induced with heat shock could overexpress diversified genes, including three whose products are deleterious to the host cells, more stably and profitably than following the log phase induction protocol. Physical and patch-clamp assays indicated that characteristics of target proteins prepared from cultures of the two different growth phases coincide. These results not only provide a better strategy for recombinant protein preparation in E. coli, but also reveal that rapid rehabilitation from stresses and stationary phase protein overproduction are fundamental characters of E. coli.
Ou
Jingxing
J
Key Laboratory of Genetic Engineering of Ministry of Education, Department of Biochemistry, College of Life Sciences, Sun Yat-Sen (Zhongshan) University, Guangzhou, People's Republic of China.
Wang
Lei
L
Ding
Xiaolan
X
Du
Jianyang
J
Zhang
Yuan
Y
Chen
Huiping
H
Xu
Anlong
A
D002455:Cell Division / Q000502:physiology; D003064:Cnidarian Venoms / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D018869:Heat-Shock Response / Q000502:physiology; D009498:Neurotoxins / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
D003064:Cnidarian Venoms; D009498:Neurotoxins; D011994:Recombinant Proteins