High-Level Gene Expression for Recombinant Penicillin Acylase Production Using the araB Promoter System in Escherichia coli

2006 ◽  
Vol 22 (6) ◽  
pp. 1518-1523 ◽  
Author(s):  
N. Narayanan ◽  
Y. Xu ◽  
C.P. Chou
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Penicillin Acylase ◽  
High Level ◽  
Promoter System

Characterization of the T7 promoter system for expressing penicillin acylase in Escherichia coli

2006 ◽  
Vol 72 (3) ◽  
pp. 529-536 ◽  
Author(s):  
Yali Xu ◽  
Stefan Rosenkranz ◽  
Chiao-Ling Weng ◽  
Jeno M. Scharer ◽  
Murray Moo-Young ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Penicillin Acylase ◽  
T7 Promoter ◽  
Promoter System

scholarly journals First report of a tetracycline-inducible gene expression system for mollicutes

Microbiology ◽  
2010 ◽  
Vol 156 (1) ◽  
pp. 198-205 ◽  
Author(s):  
Marc Breton ◽  
Evelyne Sagné ◽  
Sybille Duret ◽  
Laure Béven ◽  
Christine Citti ◽  
...  
Keyword(s):  
Gene Expression ◽  
Expression System ◽  
Inducible Promoter ◽  
High Level Expression ◽  
Spiroplasma Citri ◽  
High Level ◽  
Tetr Repressor ◽  
Promoter System ◽  
Inducible Gene

Inducible promoter systems are powerful tools for studying gene function in prokaryotes but have never been shown to function in mollicutes. In this study we evaluated the efficacy of the tetracycline-inducible promoter Pxyl/tetO2 from Bacillus subtilis in controlling gene expression in two mollicutes, the plant pathogen Spiroplasma citri and the animal pathogen Mycoplasma agalactiae. An S. citri plasmid carrying the spiralin gene under the control of the xyl/tetO2 tetracycline-inducible promoter and the TetR repressor gene under the control of a constitutive spiroplasmal promoter was introduced into the spiralin-less S. citri mutant GII3-9a3. In the absence of tetracycline, expression of TetR almost completely abolished expression of spiralin from the xyl/tetO2 promoter. Adding tetracycline (>50 ng ml−1) to the medium induced high-level expression of spiralin. Interestingly, inducible expression of spiralin was also detected in vivo: in S. citri-infected leafhoppers fed on tetracycline-containing medium and in S. citri-infected plants watered with tetracycline. A similar construct was introduced into the M. agalactiae chromosome through transposition. Tetracycline-induced expression of spiralin proved the TetR-Pxyl/tetO2 system to be functional in the ruminant pathogen, suggesting that this tetracycline-inducible promoter system might be of general use in mollicutes.

scholarly journals The Escherichia colirhaSR-PrhaBADInducible Promoter System Allows Tightly Controlled Gene Expression over a Wide Range in Pseudomonas aeruginosa

2016 ◽  
Vol 82 (22) ◽  
pp. 6715-6727 ◽  
Author(s):  
Jeffrey Meisner ◽  
Joanna B. Goldberg
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Pseudomonas Aeruginosa ◽  
Catabolite Repression ◽  
Inducible Promoter ◽  
Translational Initiation ◽  
Content Type ◽  
E Coli ◽  
Wide Range ◽  
Promoter System

ABSTRACTThearaC-ParaBADinducible promoter system is tightly controlled and allows gene expression to be modulated over a wide range inEscherichia coli, which has led to its widespread use in other bacteria. Although anecdotal evidence suggests thataraC-ParaBADis leaky inPseudomonas aeruginosa, neither a thorough analysis of this inducible promoter system inP. aeruginosanor a concerted effort to identify alternatives with improved functionality has been reported. Here, we evaluated the functionality of thearaC-ParaBADsystem inP. aeruginosa. Using transcriptional fusions to alacZreporter gene, we determined that the noninduced expression fromaraC-ParaBADis high and cannot be reduced by carbon catabolite repression as it can inE. coli. Modulating translational initiation by altering ribosome-binding site strength reduced the noninduced activity but also decreased the maximal induced activity and narrowed the induction range. Integrating the inducible promoter system into the posttranscriptional regulatory network that controls catabolite repression inP. aeruginosasignificantly decreased the noninduced activity and increased the induction range. In addition to these improvements in the functionality of thearaC-ParaBADsystem, we found that thelacIq-PtacandrhaSR-PrhaBADinducible promoter systems had significantly lower noninduced expression and were inducible over a broader range thanaraC-ParaBAD. We demonstrated that noninduced expression from thearaC-ParaBADsystem supported the function of genes involved in antibiotic resistance and tryptophan biosynthesis inP. aeruginosa, problems that were avoided withrhaSR-PrhaBAD. rhaSR-PrhaBADis tightly controlled, allows gene expression over a wide range, and represents a significant improvement overaraC-ParaBADinP. aeruginosa.IMPORTANCEWe report the shortcomings of the commonly usedEscherichia coli araC-ParaBADinducible promoter system inPseudomonas aeruginosa, successfully reengineered it to improve its functionality, and show that theE. colirhaSR-PrhaBADsystem is tightly controlled and allows inducible gene expression over a wide range inP. aeruginosa.

scholarly journals Global Change of Gene Expression and Cell Physiology in YidC-Depleted Escherichia coli

2010 ◽  
Vol 192 (8) ◽  
pp. 2193-2209 ◽  
Author(s):  
Peng Wang ◽  
Andreas Kuhn ◽  
Ross E. Dalbey
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Stress Protein ◽  
Gene Chip ◽  
Cell Physiology ◽  
Global Changes ◽  
Protein Insertion ◽  
Genes Encoding ◽  
Membrane Protein Insertion ◽  
High Level

ABSTRACT YidC depletion affects membrane protein insertion and leads to a defect in the growth of the Escherichia coli cell. We analyzed global changes in gene expression upon YidC depletion to determine the importance of YidC for cellular functions using a gene chip method to compare the transcriptomes of JS71 (control) and JS7131 (yidC depletion strain). Of the more than 4,300 genes identified, 163 were upregulated and 99 were downregulated upon YidC depletion, including genes which are responsible for DNA/RNA repair; energy metabolism; various transporters, proteases and chaperones; stress response; and translation and transcription functions. Real-time PCR was performed on selected genes to confirm the results. Specifically, we found upregulation of the genes encoding the energy transduction proteins F1Fo ATP synthase and cytochrome bo 3 oxidase due to perturbation in assembly when YidC was depleted. We also determined that the high-level induction of the PspA stress protein under YidC depletion conditions is roughly 10-fold higher than the activation due to the addition of protonophore carbonyl cyanide m-chlorophenylhydrazone (CCCP), which dissipates the proton motive force. In addition, the gene chip data reveal the Cpx stress pathway is activated upon YidC depletion. The data show the broad physiological contribution of YidC to the bacterial cell and the considerable ramification to the cell when it is depleted.

scholarly journals Genetic construction and high-level gene expression in Escherichia coli of a precursor of salmon calcitonin I.

1988 ◽  
Vol 52 (11) ◽  
pp. 2823-2830 ◽  
Author(s):  
Muneki OHMORI ◽  
Hiroyuki NARUSHIMA ◽  
Tetsuzo MIKI ◽  
Takaomi IKARI ◽  
Naganori NUMAO ◽  
...  
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Salmon Calcitonin ◽  
Expression In Escherichia Coli ◽  
High Level

Gene Expression Differences in White Blood Cells after Escherichia coli Infection in Chickens

2012 ◽  
Author(s):  
Erin Sandford ◽  
Megan Orr ◽  
Xianyao Li ◽  
Huaijun Zhou ◽  
timothy J. Johnson ◽  
...  
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Blood Cells ◽  
White Blood Cells ◽  
Escherichia Coli Infection
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