scholarly journals High-yield production, purification and characterization of functional human duodenal cytochrome b in an Escherichia coli system

2011 ◽  
Vol 79 (1) ◽  
pp. 115-121 ◽  
Author(s):  
Wen Liu ◽  
Gang Wu ◽  
Ah-Lim Tsai ◽  
Richard J. Kulmacz
Keyword(s):  
Escherichia Coli ◽  
Cytochrome B ◽  
High Yield ◽  
Purification And Characterization ◽  
Yield Production ◽  
High Yield Production ◽  
Duodenal Cytochrome B

scholarly journals High-yield production of bacteriorhodopsin via expression of a synthetic gene in Escherichia coli

1993 ◽  
Vol 211 (1-2) ◽  
pp. 27-35 ◽  
Author(s):  
Markus POMPEJUS ◽  
Karlheinz FRIEDRICH ◽  
Michael TEUFEL ◽  
Hans-Joachim FRITZ
Keyword(s):  
Escherichia Coli ◽  
Synthetic Gene ◽  
High Yield ◽  
Yield Production ◽  
High Yield Production

scholarly journals Structure-based Design of Prefusion-stabilized SARS-CoV-2 Spikes

2020 ◽  
Author(s):  
Ching-Lin Hsieh ◽  
Jory A. Goldsmith ◽  
Jeffrey M. Schaub ◽  
Andrea M. DiVenere ◽  
Hung-Che Kuo ◽  
...  
Keyword(s):  
Room Temperature ◽  
Protein A ◽  
Public Health Emergency ◽  
High Yield ◽  
The Novel ◽  
Yield Production ◽  
Novel Coronavirus ◽  
Serological Diagnostics ◽  
High Yield Production

ABSTRACTThe COVID-19 pandemic caused by the novel coronavirus SARS-CoV-2 has led to accelerated efforts to develop therapeutics, diagnostics, and vaccines to mitigate this public health emergency. A key target of these efforts is the spike (S) protein, a large trimeric class I fusion protein that is metastable and difficult to produce recombinantly in large quantities. Here, we designed and expressed over 100 structure-guided spike variants based upon a previously determined cryo-EM structure of the prefusion SARS-CoV-2 spike. Biochemical, biophysical and structural characterization of these variants identified numerous individual substitutions that increased protein yields and stability. The best variant, HexaPro, has six beneficial proline substitutions leading to ∼10-fold higher expression than its parental construct and is able to withstand heat stress, storage at room temperature, and multiple freeze-thaws. A 3.2 Å-resolution cryo-EM structure of HexaPro confirmed that it retains the prefusion spike conformation. High-yield production of a stabilized prefusion spike protein will accelerate the development of vaccines and serological diagnostics for SARS-CoV-2.

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