Construction and Characterization of a Gradient Strength Promoter Library for Fine-Tuned Gene Expression in Bacillus licheniformis

2021 ◽  
Vol 10 (9) ◽  
pp. 2331-2339
Author(s):  
Yi Rao ◽  
Peifen Li ◽  
Xinxin Xie ◽  
Jiemin Li ◽  
Yongqing Liao ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bacillus Licheniformis ◽  
Gradient Strength ◽  
Promoter Library

scholarly journals GAPPromoter Library for Fine-Tuning of Gene Expression in Pichia pastoris

2011 ◽  
Vol 77 (11) ◽  
pp. 3600-3608 ◽  
Author(s):  
Xiulin Qin ◽  
Jiangchao Qian ◽  
Gaofeng Yao ◽  
Yingping Zhuang ◽  
Siliang Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Pichia Pastoris ◽  
Fine Tuning ◽  
Promoter Strength ◽  
Control Of Gene Expression ◽  
Content Type ◽  
Expression Levels ◽  
Precise Control ◽  
Promoter Library

ABSTRACTA library of engineered promoters of various strengths is a useful genetic tool that enables the fine-tuning and precise control of gene expression across a continuum of broad expression levels. The methylotrophic yeastPichia pastorisis a well-established expression host with a large academic and industrial user base. To facilitate manipulation of gene expression spanning a wide dynamic range inP. pastoris, we created a functional promoter library through mutagenesis of the constitutiveGAPpromoter. Using yeast-enhanced green fluorescent protein (yEGFP) as the reporter, 33 mutants were chosen to form the functional promoter library. The 33 mutants spanned an activity range between ∼0.6% and 19.6-fold of the wild-type promoter activity with an almost linear fluorescence intensity distribution. After an extensive characterization of the library, the broader applicability of the results obtained with the yEGFP reporter was confirmed using two additional reporters (β-galactosidase and methionine adenosyltransferase [MAT]) at the transcription and enzyme activity levels. Furthermore, the utility of the promoter library was tested by investigating the influence of heterologous MAT gene expression levels on cell growth andS-adenosylmethionine (SAM) production. The extensive characterization of the promoter strength enabled identification of the optimal MAT activity (around 1.05 U/mg of protein) to obtain maximal volumetric SAM production. The promoter library permits precise control of gene expression and quantitative assessment that correlates gene expression level with physiologic parameters. Thus, it is a useful toolbox for both basic and applied research inP. pastoris.

Gene expression and molecular characterization of a chaperone protein HtpG from Bacillus licheniformis

2016 ◽  
Vol 85 ◽  
pp. 179-191 ◽  
Author(s):  
Hui-Fen Lo ◽  
Bo-En Chen ◽  
Min-Guan Lin ◽  
Meng-Chun Chi ◽  
Tzu-Fan Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Molecular Characterization ◽  
Bacillus Licheniformis ◽  
Chaperone Protein

Construction, Model-Based Analysis, and Characterization of a Promoter Library for Fine-Tuned Gene Expression inBacillus subtilis

2018 ◽  
Vol 7 (7) ◽  
pp. 1785-1797 ◽  
Author(s):  
Dingyu Liu ◽  
Zhitao Mao ◽  
Jiaxin Guo ◽  
Leyi Wei ◽  
Hongwu Ma ◽  
...  
Keyword(s):  
Gene Expression ◽  
Model Based ◽  
Promoter Library ◽  
Construction Model ◽  
Model Based Analysis

scholarly journals Harnessing a Transient Gene Expression System in Nicotiana benthamiana to Explore Plant Agrochemical Transporters

Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 524
Author(s):  
Bingqi Wu ◽  
Zhiting Chen ◽  
Xiaohui Xu ◽  
Ronghua Chen ◽  
Siwei Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transient Expression ◽  
Nicotiana Benthamiana ◽  
Heterologous Gene Expression ◽  
Expression System ◽  
Functional Characterization ◽  
Transient Gene Expression ◽  
High Mobility ◽  
Gene Expression System

Functional characterization of plant agrichemical transporters provided an opportunity to discover molecules that have a high mobility in plants and have the potential to increase the amount of pesticides reaching damage sites. Agrobacterium-mediated transient expression in tobacco is simple and fast, and its protein expression efficiency is high; this system is generally used to mediate heterologous gene expression. In this article, transient expression of tobacco nicotine uptake permease (NtNUP1) and rice polyamine uptake transporter 1 (OsPUT1) in Nicotiana benthamiana was performed to investigate whether this system is useful as a platform for studying the interactions between plant transporters and pesticides. The results showed that NtNUP1 increases nicotine uptake in N. benthamiana foliar discs and protoplasts, indicating that this transient gene expression system is feasible for studying gene function. Moreover, yeast expression of OsPUT1 apparently increases methomyl uptake. Overall, this method of constructing a transient gene expression system is useful for improving the efficiency of analyzing the functions of plant heterologous transporter-encoding genes and revealed that this system can be further used to study the functions of transporters and pesticides, especially their interactions.

scholarly journals Analysis of the Mechanism by Which Glucose Inhibits Maltose Induction of MAL Gene Expression in Saccharomyces

Genetics ◽  
2000 ◽  
Vol 154 (1) ◽  
pp. 121-132
Author(s):  
Zhen Hu ◽  
Yingzi Yue ◽  
Hua Jiang ◽  
Bin Zhang ◽  
Peter W Sherwood ◽  
...  
Keyword(s):  
Gene Expression ◽  
Glucose Repression ◽  
Protein A ◽  
The Other ◽  
Maltose Permease ◽  
Inducer Exclusion ◽  
Glucose Inhibition ◽  
Independent Mechanism ◽  
Mal Genes

Abstract Expression of the MAL genes required for maltose fermentation in Saccharomyces cerevisiae is induced by maltose and repressed by glucose. Maltose-inducible regulation requires maltose permease and the MAL-activator protein, a DNA-binding transcription factor encoded by MAL63 and its homologues at the other MAL loci. Previously, we showed that the Mig1 repressor mediates glucose repression of MAL gene expression. Glucose also blocks MAL-activator-mediated maltose induction through a Mig1p-independent mechanism that we refer to as glucose inhibition. Here we report the characterization of this process. Our results indicate that glucose inhibition is also Mig2p independent. Moreover, we show that neither overexpression of the MAL-activator nor elimination of inducer exclusion is sufficient to relieve glucose inhibition, suggesting that glucose acts to inhibit induction by affecting maltose sensing and/or signaling. The glucose inhibition pathway requires HXK2, REG1, and GSF1 and appears to overlap upstream with the glucose repression pathway. The likely target of glucose inhibition is Snf1 protein kinase. Evidence is presented indicating that, in addition to its role in the inactivation of Mig1p, Snf1p is required post-transcriptionally for the synthesis of maltose permease whose function is essential for maltose induction.

scholarly journals Computational design and experimental characterization of a photo-controlled mRNA-cap guanine-N7 methyltransferase

2021 ◽  
Author(s):  
Dennis Reichert ◽  
Helena Schepers ◽  
Julian Simke ◽  
Horst Lechner ◽  
Wolfgang Dörner ◽  
...  
Keyword(s):  
Gene Expression ◽  
Computational Design ◽  
Eukaryotic Cells ◽  
Transcriptional Level ◽  
Experimental Characterization ◽  
Temporal Control ◽  
Control Of Gene Expression ◽  
Multicellular Organisms

The spatial and temporal control of gene expression at the post-transcriptional level is essential in eukaryotic cells and developing multicellular organisms. In recent years optochemical and optogenetic tools have enabled...

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