Simultaneous Expression of Multiple Proteins Under a Single Promoter inCaenorhabditis elegansvia a Versatile 2A-Based Toolkit

At least one of the procyclic acidic repetitive protein (PARP or procyclin) loci of Trypanosoma brucei is a small (5- to 6-kilobase) polycistronic transcription unit which is transcribed in an alpha-amanitin-resistant manner. Its single promoter, as mapped by run-on transcription analysis and UV inactivation of transcription, is located immediately upstream of the first alpha-PARP gene. Transcription termination occurs in a region approximately 3 kilobases downstream of the beta-PARP gene. The location of the promoter was confirmed by its ability to direct transcription of the bacterial chloramphenicol acetyltransferase gene in insect-form (procyclic) T. brucei. The putative PARP promoter is located in the region between the 3' splice acceptor site (nucleotide position 0) and nucleotide position -196 upstream of the alpha-PARP genes. Regulatory regions influencing the levels of PARP expression may be located further upstream. We conclude that a single promoter, which is located very close to the 3' splice acceptor site of the alpha-PARP genes, directs the transcription of a small, polycistronic, and alpha-amanitin-resistant transcription unit.

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The simultaneous expression of both ephrin B3 receptor and E-cadherin in Barrett`s adenocarcinoma is associated with favorable clinical staging

European Journal of Medical Research ◽

10.1186/2047-783x-17-10 ◽

2012 ◽

Vol 17 (1) ◽

pp. 10 ◽

Cited By ~ 7

Author(s):

Matthias C Schauer ◽

Nikolas H Stoecklein ◽

Joerg Theisen ◽

Feride Kröpil ◽

Stephan Baldus ◽

...

Keyword(s):

Clinical Staging ◽

Simultaneous Expression ◽

E Cadherin

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Regulation of the Bacillus subtilis bcrC Bacitracin Resistance Gene by Two Extracytoplasmic Function σ Factors

Journal of Bacteriology ◽

10.1128/jb.184.22.6123-6129.2002 ◽

2002 ◽

Vol 184 (22) ◽

pp. 6123-6129 ◽

Cited By ~ 91

Author(s):

Min Cao ◽

John D. Helmann

Keyword(s):

Bacillus Subtilis ◽

Resistance Gene ◽

Mutant Strain ◽

Stress Responses ◽

Double Mutant ◽

De Novo ◽

De Novo Synthesis ◽

Gene Encoding ◽

Single Promoter ◽

Higher Sensitivity

ABSTRACT Bacitracin resistance is normally conferred by either of two major mechanisms, the BcrABC transporter, which pumps out bacitracin, or BacA, an undecaprenol kinase that provides C55-isoprenyl phosphate by de novo synthesis. We demonstrate that the Bacillus subtilis bcrC (ywoA) gene, encoding a putative bacitracin transport permease, is an important bacitracin resistance determinant. A bcrC mutant strain had an eightfold-higher sensitivity to bacitracin. Expression of bcrC initiated from a single promoter site that could be recognized by either of two extracytoplasmic function (ECF) σ factors, σX or σM. Bacitracin induced expression of bcrC, and this induction was dependent on σM but not on σX. Under inducing conditions, expression was primarily dependent on σM. As a consequence, a sigM mutant was fourfold more sensitive to bacitracin, while the sigX mutant was only slightly sensitive. A sigX sigM double mutant was similar to a bcrC mutant in sensitivity. These results support the suggestion that one function of B. subtilis ECF σ factors is to coordinate antibiotic stress responses.

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A single promoter system co-expressing RNA sensor with fluorescent proteins for quantitative mRNA imaging in living tumor cells

Chemical Science ◽

10.1039/c9sc00458k ◽

2019 ◽

Vol 10 (18) ◽

pp. 4828-4833 ◽

Cited By ~ 8

Author(s):

Zhan-Ming Ying ◽

Yue-Yan Yuan ◽

Bin Tu ◽

Li-Juan Tang ◽

Ru-Qin Yu ◽

...

Keyword(s):

Tumor Cells ◽

Live Cell Imaging ◽

Cell Imaging ◽

Fluorescent Proteins ◽

Live Cell ◽

Rna Aptamers ◽

Rna Sensors ◽

Promoter System ◽

Single Promoter

Genetically encoded light-up RNA aptamers afford a valuable platform for developing RNA sensors toward live cell imaging.

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Divergent MLS1 promoters lie on a fitness plateau for gene expression

10.1101/026427 ◽

2015 ◽

Author(s):

Andrew C Bergen ◽

Gerilyn M Olsen ◽

Justin C Fay

Keyword(s):

Gene Expression ◽

Saccharomyces Cerevisiae ◽

Dynamic Response ◽

Yeast Species ◽

Glucose Repression ◽

Gene Activation ◽

Malate Synthase ◽

Expression Divergence ◽

Fitness Effects ◽

Single Promoter

Qualitative patterns of gene activation and repression are often conserved despite an abundance of quantitative variation in expression levels within and between species. A major challenge to interpreting patterns of expression divergence is knowing which changes in gene expression affect fitness. To characterize the fitness effects of gene expression divergence we placed orthologous promoters from eight yeast species upstream of malate synthase (MLS1) in Saccharomyces cerevisiae. As expected, we found these promoters varied in their expression level under activated and repressed conditions as well as in their dynamic response following loss of glucose repression. Despite these differences, only a single promoter driving near basal levels of expression caused a detectable loss of fitness. We conclude that the MLS1 promoter lies on a fitness plateau whereby even large changes in gene expression can be tolerated without a substantial loss of fitness.

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