High level soluble expression, purification, and characterization of human ciliary neuronotrophic factor in Escherichia coli by single protein production system

2014 ◽  
Vol 96 ◽  
pp. 8-13 ◽  
Author(s):  
Ke Wang ◽  
Fanfan Zhou ◽  
Lan Zhu ◽  
Xue Zhu ◽  
Kai Zhang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Production System ◽  
Protein Production ◽  
Soluble Expression ◽  
Purification And Characterization ◽  
Single Protein ◽  
High Level ◽  
Ciliary Neuronotrophic Factor

High-level soluble expression, purification and characterization of active human midkine from Escherichia coli

2010 ◽  
Vol 70 (2) ◽  
pp. 270-276 ◽  
Author(s):  
W. Kelly Yan ◽  
Marjo Goette ◽  
Gabriele Hofmann ◽  
Isabel Zaror ◽  
Janet Sim
Keyword(s):  
Escherichia Coli ◽  
Soluble Expression ◽  
Purification And Characterization ◽  
High Level

scholarly journals Secretion of human superoxide dismutase in Escherichia coli using the condensed single-protein-production system

2010 ◽  
Vol 19 (12) ◽  
pp. 2330-2335 ◽  
Author(s):  
Lili Mao ◽  
Peter B. Stathopulos ◽  
Mitsuhiko Ikura ◽  
Masayori Inouye
Keyword(s):  
Escherichia Coli ◽  
Superoxide Dismutase ◽  
Production System ◽  
Protein Production ◽  
Single Protein

scholarly journals Use of Amino Acids as Inducers for High-Level Protein Expression in the Single-Protein Production System

2010 ◽  
Vol 76 (18) ◽  
pp. 6063-6068 ◽  
Author(s):  
S. Thangminlal Vaiphei ◽  
Lili Mao ◽  
Tsutomu Shimazu ◽  
Jung-Ho Park ◽  
Masayori Inouye
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Protein Production ◽  
Target Protein ◽  
Complete Suppression ◽  
Tryptophan Residues ◽  
Single Protein ◽  
High Level ◽  
Isotopic Incorporation ◽  
Specific Mrna

ABSTRACT By taking advantage of MazF, an ACA codon-specific mRNA interferase, Escherichia coli cells can be converted into a bioreactor producing only a single protein of interest by using an ACA-less mRNA for the protein. In this single-protein production (SPP) system, we engineered MazF by replacing two tryptophan residues in positions 14 and 83 with Phe (W14F) and Leu (W83L), respectively. Upon the addition of an inducer (IPTG [isopropyl-β-d-thiogalactopyranoside]), the mutated MazF [MazF(ΔW)] can still be produced even in the absence of tryptophan in the medium by using a Trp auxotroph, while a target protein having Trp residues cannot be produced. However, at 3 h after the addition of IPTG, the addition of tryptophan to the medium exclusively induces production of the target protein at a high level. A similar SPP system was also constructed with the use of a His-less protein [MazF(ΔH)] and a His auxotroph. Using these dual-induction systems, isotopic enrichments of 13C, 15N, and 2H were highly improved by almost complete suppression of the production of the unlabeled target protein. In both systems, isotopic incorporation reached more than 98% labeling efficiency, significantly reducing the background attributable to the unlabeled target protein.

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