scholarly journals High-Throughput Beta-glucuronidase (GUS) assay for Phaeodactylum tricornutum v1 (protocols.io.bbexijfn)

protocols.io ◽  
2020 ◽  
Author(s):  
Erin Garza ◽  
Vincent Bielinski
Keyword(s):  
High Throughput ◽  
Phaeodactylum Tricornutum ◽  
Gus Assay

scholarly journals Chemical Mutagenesis and Fluorescence-Based High-Throughput Screening for Enhanced Accumulation of Carotenoids in a Model Marine Diatom Phaeodactylum tricornutum

Marine Drugs ◽  
2018 ◽  
Vol 16 (8) ◽  
pp. 272 ◽  
Author(s):  
Zhiqian Yi ◽  
Yixi Su ◽  
Maonian Xu ◽  
Andreas Bergmann ◽  
Saevar Ingthorsson ◽  
...  
Keyword(s):  
High Throughput ◽  
Metabolic Pathways ◽  
Phaeodactylum Tricornutum ◽  
Mass Spectrometry Data ◽  
Marine Diatom ◽  
Chemical Mutagenesis ◽  
Carotenoid Content ◽  
Enzymatic Reactions ◽  
Wild Type ◽  
A Genome

Diatoms are a major group of unicellular algae that are rich in lipids and carotenoids. However, sustained research efforts are needed to improve the strain performance for high product yields towards commercialization. In this study, we generated a number of mutants of the model diatom Phaeodactylum tricornutum, a cosmopolitan species that has also been found in Nordic region, using the chemical mutagens ethyl methanesulfonate (EMS) and N-methyl-N′-nitro-N-nitrosoguanidine (NTG). We found that both chlorophyll a and neutral lipids had a significant correlation with carotenoid content and these correlations were better during exponential growth than in the stationary growth phase. Then, we studied P. tricornutum common metabolic pathways and analyzed correlated enzymatic reactions between fucoxanthin synthesis and pigmentation or lipid metabolism through a genome-scale metabolic model. The integration of the computational results with liquid chromatography-mass spectrometry data revealed key compounds underlying the correlative metabolic pathways. Approximately 1000 strains were screened using fluorescence-based high-throughput method and five mutants selected had 33% or higher total carotenoids than the wild type, in which four strains remained stable in the long term and the top mutant exhibited an increase of 69.3% in fucoxanthin content compared to the wild type. The platform described in this study may be applied to the screening of other high performing diatom strains for industrial applications.

Application of the GUS marker gene technique to high-throughput screening of rhizobial competition

1999 ◽  
Vol 45 (8) ◽  
pp. 678-685 ◽  
Author(s):  
Kate J Wilson ◽  
Adriana Parra ◽  
Lina Botero
Keyword(s):  
Phaseolus Vulgaris ◽  
Sample Size ◽  
Microbial Ecology ◽  
High Throughput ◽  
High Throughput Screening ◽  
Marker Gene ◽  
Nodule Occupancy ◽  
Gus Gene ◽  
Gus Assay ◽  
Rhizobium Strains

The GUS marker gene system has been developed for the study of bacterial ecology, particularly rhizobial competition. For high-throughput field screening of rhizobial competition, the technique must be robust and reliable under diverse conditions, with diverse cultivars and strains. Here we demonstrate its applicability to the evaluation of competition on five different Phaseolus vulgaris cultivars with 10 different Rhizobium strains. We describe refinements of the GUS assay, which make it more affordable and applicable to field-based studies, and use the assay to examine the effect of sample size on the accuracy of nodule occupancy measurements.Key words: GUS gene, Rhizobium, rhizobial competition, microbial ecology.

154: Integration of TPSA and High-Throughput Mass Spectrometry Data Improves Prostate Cancer Prediction

2007 ◽  
Vol 177 (4S) ◽  
pp. 52-53
Author(s):  
Stefano Ongarello ◽  
Eberhard Steiner ◽  
Regina Achleitner ◽  
Isabel Feuerstein ◽  
Birgit Stenzel ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Mass Spectrometry ◽  
High Throughput ◽  
Mass Spectrometry Data ◽  
Cancer Prediction
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