Using the pER8:GUS Reporter System to Screen for Phytoestrogens fromCaesalpinia sappan

2011 ◽  
Vol 74 (8) ◽  
pp. 1698-1706 ◽  
Author(s):  
Wan-Chun Lai ◽  
Hui-Chun Wang ◽  
Guan-Yu Chen ◽  
Juan-Cheng Yang ◽  
Michal Korinek ◽  
...  
Keyword(s):  
Reporter System ◽  
Gus Reporter ◽  
Gus Reporter System

scholarly journals Bioactivity guided isolation of phytoestrogenic compounds from Cyclopia genistoides by the pER8:GUS reporter system

2017 ◽  
Vol 110 ◽  
pp. 201-207 ◽  
Author(s):  
O. Roza ◽  
W.-C. Lai ◽  
I. Zupkó ◽  
J. Hohmann ◽  
N. Jedlinszki ◽  
...  
Keyword(s):  
Reporter System ◽  
Gus Reporter ◽  
Gus Reporter System

The GUS Reporter System in Flower Development Studies

2013 ◽  
pp. 295-304 ◽  
Author(s):  
Janaki S. Mudunkothge ◽  
Beth A. Krizek
Keyword(s):  
Flower Development ◽  
Development Studies ◽  
Reporter System ◽  
Gus Reporter ◽  
Gus Reporter System

The GUS Reporter System as a Tool to Study Plant Gene Expression

Gus Protocols ◽  
1992 ◽  
pp. 23-43 ◽  
Author(s):  
Thomas Martin ◽  
Rosa-Valentina Wöhner ◽  
Sabine Hummel ◽  
Lothar Willmitzer ◽  
Wolf B. Frommer
Keyword(s):  
Gene Expression ◽  
Reporter System ◽  
Plant Gene Expression ◽  
Plant Gene ◽  
Gus Reporter ◽  
Gus Reporter System

Investigation of the phytoestrogenic activity of Cyclopia genistoides by pER8:GUS reporter system

Planta Medica ◽  
2014 ◽  
Vol 80 (16) ◽  
Author(s):  
O Roza ◽  
WC Lai ◽  
FR Chang ◽  
D Csupor ◽  
K Boros
Keyword(s):  
Reporter System ◽  
Gus Reporter ◽  
Gus Reporter System

scholarly journals Chromosomal Location Plays a Role in Regulation of Aflatoxin Gene Expression in Aspergillus parasiticus

2002 ◽  
Vol 68 (1) ◽  
pp. 306-315 ◽  
Author(s):  
Ching-Hsun Chiou ◽  
Michael Miller ◽  
David L. Wilson ◽  
Frances Trail ◽  
John E. Linz
Keyword(s):  
Gene Cluster ◽  
Promoter Activity ◽  
Aspergillus Parasiticus ◽  
Chromosomal Location ◽  
Selectable Marker ◽  
Growth Conditions ◽  
Gus Expression ◽  
Reporter System ◽  
Gus Reporter ◽  
Homologous Site

ABSTRACT The nor-1 gene in the filamentous fungus Aspergillus parasiticus encodes a ketoreductase involved in aflatoxin biosynthesis. To study environmental influences on nor-1 expression, we generated plasmid pAPGUSNNB containing a nor-1 promoter-β-glucuronidase (GUS) (encoded by uidA) reporter fusion with niaD (encodes nitrate reductase) as a selectable marker. niaD transformants of A. parasiticus strain NR-1 (niaD) carried pAPGUSNNB integrated predominantly at the nor-1 or niaD locus. Expression of the native nor-1 and nor-1::GUS reporter was compared in transformants grown under aflatoxin-inducing conditions by Northern and Western analyses and by qualitative and quantitative GUS activity assays. The timing and level of nor-1 promoter function with pAPGUSNNB integrated at nor-1 was similar to that observed for the native nor-1 gene. In contrast, nor-1 promoter activity in pAPGUSNNB and a second nor-1::GUS reporter construct, pBNG3.0, was not detectable when integration occurred at niaD. Because niaD-dependent regulation could account for the absence of expression at niaD, a third chromosomal location was analyzed using pAPGUSNP, which contained nor-1::GUS plus pyrG (encodes OMP decarboxylase) as a selectable marker. GUS expression was detectable only when pAPGUSNP integrated at nor-1 and was not detectable at pyrG, even under growth conditions that required pyrG expression. nor-1::GUS is regulated similarly to the native nor-1 gene when it is integrated at its homologous site within the aflatoxin gene cluster but is not expressed at native nor-1 levels at two locations outside of the aflatoxin gene cluster. We conclude that the GUS reporter system can be used effectively to measure nor-1 promoter activity and that nor-1 is subject to position-dependent regulation in the A. parasiticus chromosome.

Length, orientation, and plant host influence the mutation frequency in microsatellites

Genome ◽  
2006 ◽  
Vol 49 (11) ◽  
pp. 1366-1373 ◽  
Author(s):  
Aïda Azaiez ◽  
Éric F. Bouchard ◽  
Martine Jean ◽  
François J. Belzile
Keyword(s):  
Mutation Frequency ◽  
Dna Repeats ◽  
Reporter Construct ◽  
Reporter System ◽  
Plant Host ◽  
Factors Affecting ◽  
Gus Reporter ◽  
Somatic Tissues ◽  
Host Influence ◽  
Polymerase Slippage

Microsatellites are simple, tandem DNA repeats that represent unstable regions of the genome. They undergo frequent changes in tract length by base additions or deletions due to DNA polymerase slippage during replication. To characterize factors affecting the frequency of spontaneous mutations occurring in microsatellites in plants, a reporter system was used in Arabidopsis thaliana and tomato ( Lycopersicon esculentum ). The β-glucuronidase (GUS) reporter system was used to measure the mutation frequency in various microsatellites (G7, G10, G13, G16, and C16) in somatic tissues. Our results indicate that this frequency increases with the number of repeats: a G16 tract was almost 80-fold more mutable than a G7 tract. Furthermore, the frequency of mutations depends on repeat orientation, as G16 was 3-fold more mutable than C16. The mutation rate was also found to differ markedly in Arabidopsis and tomato for an identical microsatellite. Indeed, Arabidopsis showed a 5-fold higher mutation frequency than tomato with the same G7 reporter construct. Finally, mutation in a G16 tract was frequent enough that mutations transmitted germinally to the next generation could be detected at a relatively high frequency.

scholarly journals Transformation of European Ash (Fraxinus excelsior L.) Callus as a Starting Point for Understanding the Molecular Basis of Ash Dieback

Plants ◽  
2021 ◽  
Vol 10 (11) ◽  
pp. 2524
Author(s):  
Anna Hebda ◽  
Aleksandra Liszka ◽  
Piotr Zgłobicki ◽  
Katarzyna Nawrot-Chorabik ◽  
Jan J. Lyczakowski
Keyword(s):  
Molecular Basis ◽  
Selection Process ◽  
Pathogenic Fungus ◽  
Fraxinus Excelsior ◽  
Reporter System ◽  
Ash Dieback ◽  
Gus Reporter ◽  
Hymenoscyphus Fraxineus ◽  
Starting Point ◽  
European Ash

The population of European ash (Fraxinus excelsior L.) is currently facing the risk of collapse, mainly due to ash dieback, a disease caused by a pathogenic fungus, Hymenoscyphus fraxineus. To facilitate studies into the molecular basis of ash dieback and design breeding strategies for a generation of resistant trees, it is necessary to develop tools enabling the study of gene function in F. excelsior. Despite this, a method for the genetic engineering of F. excelsior is still missing. Here, we report the first successful genetic transformation of F. excelsior callus and a selection process enabling the formation of stable transgenic callus lines. The protocol relies on the use of Agrobacterium tumefaciens to transform callus tissue derived from embryos of F. excelsior. In our experiments, we used the β-glucuronidase (GUS) reporter system to demonstrate the transformation of callus cells and performed RT-PCR experiments to confirm the stable expression of the transgene. Since ash dieback threatens the long-term stability of many native F. excelsior populations, we hope that the transformation techniques described in this manuscript will facilitate rapid progress in uncovering the molecular basis of the disease and the validation of gene targets previously proposed to be linked to the resistance of trees to H. fraxineus pathogenicity.

scholarly journals Transformation of European ash (Fraxinus excelsior L.) callus as a starting point for understanding the molecular basis of ash dieback.

2020 ◽  
Author(s):  
Anna Hebda ◽  
Aleksandra Liszka ◽  
Piotr Zgłobicki ◽  
Katarzyna Nawrot-Chorabik ◽  
Jan J Lyczakowski
Keyword(s):  
Molecular Basis ◽  
Callus Tissue ◽  
Selection Process ◽  
Genetic Material ◽  
Fraxinus Excelsior ◽  
Reporter System ◽  
Ash Dieback ◽  
Gus Reporter ◽  
Starting Point ◽  
European Ash

Plant genetic engineering requires transfer of genetic material into plant cells, a process that is frequently challenging for tree species. Here we report first successful genetic transformation of European ash (Fraxinus excelsior). The protocol relies on the use of Agrobacterium tumefaciens to transform callus tissue derived from embryos of F. excelsior. In our experiments, we use the β-glucuronidase (GUS) reporter system to demonstrate transformation of ash callus tissue. Moreover, we describe an antibiotic-resistance based selection process enabling formation of stable transgenic callus lines. Since ash dieback threatens the long-term stability of many native F. excelsior populations, we hope that the transformation techniques described in this manuscript will facilitate rapid progress in uncovering the molecular basis of the disease and mechanisms used by trees to resist it.

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