scholarly journals Influence of various inductor concentrations on the human alpha-2b interferon production in the bacteriophage T7 RNA polymerase-base expression system in Escherichia coli cells

2003 ◽  
Vol 19 (5) ◽  
pp. 457-462 ◽  
Author(s):  
I. Yu. Slavchenko ◽  
E. V. Boreyko ◽  
N. V. Vorobey
Keyword(s):  
Escherichia Coli ◽  
Rna Polymerase ◽  
Expression System ◽  
T7 Rna Polymerase ◽  
Bacteriophage T7 ◽  
Interferon Production ◽  
Escherichia Coli Cells

scholarly journals Engineering efficient termination of bacteriophage T7 RNA polymerase transcription

2021 ◽  
Author(s):  
Diana Gabriela Calvopina Chavez ◽  
Mikaela Anne Gardner ◽  
Joel S Griffitts
Keyword(s):  
Rna Polymerase ◽  
Molecular Level ◽  
Expression System ◽  
T7 Rna Polymerase ◽  
Bacteriophage T7 ◽  
Lower Efficiency ◽  
Transcriptional Termination ◽  
T7 Polymerase

The bacteriophage T7 expression system is one of the most prominent transcription systems used in biotechnology and molecular-level research. However, T7 RNA polymerase is prone to read-through transcription due to its high processivity. As a consequence, enforcing efficient transcriptional termination is difficult. The termination hairpin found natively in the T7 genome is adapted to be inefficient, exhibiting 62% termination efficiency in vivo and even lower efficiency in vitro. In this study, we engineered a series of sequences that outperform the efficiency of the native terminator hairpin. By embedding a previously discovered 8-nucleotide T7 polymerase pause sequence within a synthetic hairpin sequence, we observed in vivo termination efficiency of 91%; by joining two short sequences into a tandem 2-hairpin structure, termination efficiency was increased to 98% in vivo and 91% in vitro. This study also tests the ability of these engineered sequences to terminate transcription of the Escherichia coli RNA polymerase. Two out of three of the most successful T7 polymerase terminators also facilitated termination of the bacterial polymerase with around 99% efficiency.

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