Development of orthogonal T7 expression system in Klebsiella pneumoniae

2020 ◽  
Vol 117 (8) ◽  
pp. 2446-2459
Author(s):  
Peng Zhao ◽  
Minrui Ren ◽  
Xizhen Ge ◽  
Pingfang Tian ◽  
Tianwei Tan
Keyword(s):  
Klebsiella Pneumoniae ◽  
Expression System ◽  
T7 Expression System

Production of HIV-1 gp120 in Packed-Bed Bioreactor Using the Vaccinia Virus/T7 Expression System

2000 ◽  
Vol 16 (5) ◽  
pp. 744-750 ◽  
Author(s):  
Y.-C. Hu ◽  
J. Kaufman ◽  
M.W. Cho ◽  
H. Golding ◽  
J. Shiloach
Keyword(s):  
Vaccinia Virus ◽  
Expression System ◽  
Packed Bed ◽  
Packed Bed Bioreactor ◽  
T7 Expression System ◽  
Hiv 1

A T7-expression system under temperature control could create temperature-sensitive phenotype of target gene in Escherichia coli

2007 ◽  
Vol 68 (3) ◽  
pp. 497-506 ◽  
Author(s):  
Rubing Liang ◽  
Xipeng Liu ◽  
Jianhua Liu ◽  
Qiushi Ren ◽  
Peiji Liang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Temperature Control ◽  
Target Gene ◽  
Expression System ◽  
Temperature Sensitive ◽  
T7 Expression System ◽  
Temperature Sensitive Phenotype

Construction of a host-independent T7 expression system with small RNA regulation

2014 ◽  
Vol 189 ◽  
pp. 72-75 ◽  
Author(s):  
Gang Wang ◽  
Qiang Li ◽  
Dikai Xu ◽  
Mingxin Cui ◽  
Xiao Sun ◽  
...  
Keyword(s):  
Small Rna ◽  
Expression System ◽  
Rna Regulation ◽  
T7 Expression System

scholarly journals A novel gene expression system for Ralstonia eutropha based on the T7 promoter

2020 ◽  
Author(s):  
Muzi Hu ◽  
Bin Xiong ◽  
Zhongkang Li ◽  
Li Liu ◽  
Siwei Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Production ◽  
Ralstonia Eutropha ◽  
Expression System ◽  
Expression Plasmid ◽  
T7 Promoter ◽  
T7 Expression System ◽  
Plasmid Size ◽  
Rna Polymerase Gene ◽  
Electroporation Efficiency

Abstract Background: Ralstonia eutropha (syn. Cupriavidus necator) is a model microorganism for studying metabolism of polyhydroxyalkanoates (PHAs) and a potential chassis for protein expression due to various advantages. Although current plasmid systems of R. eutropha provide a basic platform for gene expression, the performance of the expression-inducing systems is still limited. In addition, the sizes of the cloned genes are limited due to the large sizes of the plasmid backbones.Results: In this study, an R. eutropha T7 expression system was established by integrating a T7 RNA polymerase gene driven by the PBAD promoter into the genome of R. eutropha, as well as adding a T7 promoter into a pBBR1-derived plasmid for gene expression. In addition, the essential DNA sequence necessary for pBBR1 plasmid replication was identified, and the redundant parts were deleted reducing the expression plasmid size to 3392 bp, which improved the electroporation efficiency about 4 times. As a result, the highest expression level of RFP was enhanced, and the L-arabinose concentration for expression induction was decreased 20 times. Conclusions: The R. eutropha T7 expression system provides an efficient platform for protein production and synthetic biology applications.

scholarly journals A novel gene expression system for Ralstonia eutropha based on the T7 promoter

2020 ◽  
Author(s):  
Muzi Hu ◽  
Bin Xiong ◽  
Zhongkang Li ◽  
Li Liu ◽  
Siwei Li ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Production ◽  
Ralstonia Eutropha ◽  
Expression System ◽  
Expression Plasmid ◽  
T7 Promoter ◽  
T7 Expression System ◽  
Plasmid Size ◽  
Rna Polymerase Gene ◽  
Electroporation Efficiency

Abstract Background: Ralstonia eutropha (syn. Cupriavidus necator) is a model microorganism for studying metabolism of polyhydroxyalkanoates (PHAs) and a potential chassis for protein expression due to various advantages. Although current plasmid systems of R. eutropha provide a basic platform for gene expression, the performance of the expression-inducing systems is still limited. In addition, the sizes of the cloned genes are limited due to the large sizes of the plasmid backbones.Results: In this study, an R. eutropha T7 expression system was established by integrating a T7 RNA polymerase gene driven by the PBAD promoter into the genome of R. eutropha, as well as adding a T7 promoter into a pBBR1-derived plasmid for gene expression. In addition, the essential DNA sequence necessary for pBBR1 plasmid replication was identified, and the redundant parts were deleted reducing the expression plasmid size to 3392 bp, which improved the electroporation efficiency about 4 times. As a result, the highest expression level of RFP was enhanced, and the L-arabinose concentration for expression induction was decreased 20 times. Conclusions: The R. eutropha T7 expression system provides an efficient platform for protein production and synthetic biology applications.

CRISPR/Cas9 mediated T7 RNA polymerase gene knock-in in E. coli BW25113 makes T7 expression system work efficiently

2021 ◽  
Author(s):  
Changchuan Ye ◽  
Xi Chen ◽  
Mengjie Yang ◽  
Xiangfang Zeng ◽  
Shiyan Qiao
Keyword(s):  
Rna Polymerase ◽  
Mutant Strain ◽  
Aminolevulinic Acid ◽  
Expression System ◽  
T7 Rna Polymerase ◽  
Bacterial Strains ◽  
T7 Promoter ◽  
E Coli ◽  
T7 Expression System ◽  
High Level

Abstract T7 Expression System is a common method of ensuring tight control and high-level induced expression. However, this system can only work in some bacterial strains in which the T7 RNA Polymerase gene resides in the chromosome. In this study, we successfully introduced a chromosomal copy of the T7 RNA Polymerase gene under control of the lacUV5 promoter into Escherichia coli BW25113. The T7 Expression System worked efficiently in this mutant strain named BW25113-T7. We demonstrated that this mutant strain could satisfactorily produce 5-Aminolevulinic Acid via C5 pathway. A final study was designed to enhance the controllability of T7 Expression System in this mutant strain by constructing a T7 Promoter Variants Library. These efforts advanced E. coli BW25113-T7 to be a practical host for future metabolic engineering efforts.

Generation of an AraC-araBAD Promoter-Regulated T7 Expression System

2000 ◽  
Vol 277 (1) ◽  
pp. 67-73 ◽  
Author(s):  
Diane R. Wycuff ◽  
Kathleen S. Matthews
Keyword(s):  
Expression System ◽  
T7 Expression System

Stability studies with different vector backbones utilizing the T7 expression system inEscherichia coli

2008 ◽  
Vol 83 (8) ◽  
pp. 1120-1125 ◽  
Author(s):  
Rupali Walia ◽  
Jahar Kanti Deb ◽  
Krishna Jyoti Mukherjee
Keyword(s):  
Expression System ◽  
Inescherichia Coli ◽  
Stability Studies ◽  
T7 Expression System

scholarly journals Efficient Expression of Soluble Recombinant Protein Fused with Core-Streptavidin in Bacterial Strain with T7 Expression System

2020 ◽  
Vol 3 (4) ◽  
pp. 82
Author(s):  
Ammar Tarar ◽  
Esmael M. Alyami ◽  
Ching-An Peng
Keyword(s):  
Fusion Protein ◽  
Thymidine Phosphorylase ◽  
Expression System ◽  
Model System ◽  
Carcinoma Cells ◽  
T7 Promoter ◽  
E Coli ◽  
Efficient Expression ◽  
T7 Expression System ◽  
Soluble Recombinant Protein

The limited amount of fusion protein transported into cytosol milieu has made it challenging to obtain a sufficient amount for further applications. To avoid the laborious and expensive task, T7 promoter-driving pET-30a(+) coding for chimeric gene of thymidine phosphorylase and core streptavidin as a model system was constructed and transformed into a variety of E. coli strains with T7 expression system. Our results demonstrated that the pET-30a(+)-TP-coreSA/Lemo21(DE3) system is able to provide efficient expression of soluble TP-coreSA fusion protein for purification. Moreover, the eluted TP-coreSA fusion protein tethered on biotinylated A549 carcinoma cells could effectively eliminate these malignant cells after administrating prodrug 5′-DFUR.

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