scholarly journals Fenhexamid - an efficient and inexpensive fungicide for selection of Magnaporthe oryzae transformants

2021 ◽  
Author(s):  
Alex Wegner ◽  
Louisa Wirtz ◽  
Thomas Leisen ◽  
Matthias Hahn ◽  
Ulrich Schaffrath
Keyword(s):  
Magnaporthe Oryzae ◽  
Model Organism ◽  
Selection Procedure ◽  
Phytopathogenic Fungi ◽  
Infection Process ◽  
Ergosterol Biosynthesis ◽  
Fusarium Fujikuroi ◽  
Plant Pathogen Interactions ◽  
Efficient Selection ◽  
Selection Of

AbstractMagnaporthe oryzae is one of the most economically important phytopathogenic fungi, and is used as a model organism to study plant-pathogen interactions. To unravel the infection process, forward and reverse genetic approaches are essential, but are often hindered by the lack of a straightforward selection procedure for transformants. Here we report on the use of fenhexamid, an inhibitor of ergosterol biosynthesis, for selection of M. oryzae transformants. An allele of the sterol 3-ketoreductase gene of Fusarium fujikuroi (FfERG27), known to confer resistance to fenhexamid, has already been used successfully with transformants of Botrytis cinerea. Our results demonstrate that expression of the FfERG27 allele in M. oryzae also enables highly efficient selection of transformants on fenhexamid-containing media. The use of fenhexamid is an inexpensive alternative for selection as compared to commonly used antibiotics like hygromycin. No impact on growth and infection phenotypes of fenhexamid resistant M. oryzae mutants was detected, which underpins its usefulness for selecting M. oryzae transformants.

scholarly journals Selection of chemotaxis mutants of Dictyostelium discoideum.

1987 ◽  
Vol 104 (1) ◽  
pp. 151-161 ◽  
Author(s):  
J E Segall ◽  
P R Fisher ◽  
G Gerisch
Keyword(s):  
Dictyostelium Discoideum ◽  
Cell Population ◽  
Selection Procedure ◽  
Total Cell ◽  
Wild Type ◽  
Total Cell Population ◽  
Mutant Cells ◽  
Efficient Selection ◽  
Filter Layer ◽  
Selection Of

A method has been developed for the efficient selection of chemotaxis mutants of Dictyostelium discoideum. Mutants defective in the chemotactic response to folate could be enriched up to 30-fold in one round of selection using a chamber in which a compartment that contained the chemoattractant was separated by a sandwich of four nitrocellulose filters from a compartment that contained buffer. Mutagenized cells were placed in the center of the filter layer and exposed to the attractant gradient built up between the compartments for a period of 3-4 h. While wild-type cells moved through the filters in a wave towards the compartment that contained attractant, mutant cells remained in the filter to which they were applied. After several repetitions of the selection procedure, mutants defective in chemotaxis made up 10% of the total cell population retained in that filter. Mutants exhibiting three types of alterations were collected: motility mutants with either reduced speed of movement, or altered rates of turning; a single mutant defective in production of the attractant-degrading enzyme, folate deaminase; and mutants with normal motility but reduced chemotactic responsiveness. One mutant showed drastically reduced sensitivity in folate-induced cGMP production. Morphogenetic alterations of mutants defective in folate chemotaxis are described.

Identifying Measurement Invariant Item Sets in Cross-Cultural Settings Using an Automated Item Selection Procedure

Methodology ◽  
2018 ◽  
Vol 14 (4) ◽  
pp. 177-188 ◽  
Author(s):  
Martin Schultze ◽  
Michael Eid
Keyword(s):  
Measurement Invariance ◽  
Optimal Solution ◽  
Selection Procedure ◽  
Cross Cultural ◽  
Item Selection ◽  
Ant System ◽  
Item Quality ◽  
Ant System Algorithm ◽  
Selection For ◽  
Selection Of

Abstract. In the construction of scales intended for the use in cross-cultural studies, the selection of items needs to be guided not only by traditional criteria of item quality, but has to take information about the measurement invariance of the scale into account. We present an approach to automated item selection which depicts the process as a combinatorial optimization problem and aims at finding a scale which fulfils predefined target criteria – such as measurement invariance across cultures. The search for an optimal solution is performed using an adaptation of the [Formula: see text] Ant System algorithm. The approach is illustrated using an application to item selection for a personality scale assuming measurement invariance across multiple countries.

Combination of ribosome and phage display for fast selection of high affinity VHH antibody fragments

2019 ◽  
pp. 27-33
Author(s):  
YuE Kravchenko ◽  
SV Ivanov ◽  
DS Kravchenko ◽  
EI Frolova ◽  
SP Chumakov
Keyword(s):  
Phage Display ◽  
Selection Procedure ◽  
Free System ◽  
Phage Displays ◽  
High Affinity ◽  
Binding Domains ◽  
A Cell ◽  
Vhh Antibodies ◽  
Efficiency Of Selection ◽  
Selection Of

Selection of antibodies using phage display involves the preliminary cloning of the repertoire of sequences encoding antigen-binding domains into phagemid, which is considered the bottleneck of the method, limiting the resulting diversity of libraries and leading to the loss of poorly represented variants before the start of the selection procedure. Selection in cell-free conditions using a ribosomal display is devoid from this drawback, however is highly sensitive to PCR artifacts and the RNase contamination. The aim of the study was to test the efficiency of a combination of both methods, including pre-selection in a cell-free system to enrich the source library, followed by cloning and final selection using phage display. This approach may eliminate the shortcomings of each method and increase the efficiency of selection. For selection, alpaca VHH antibody sequences suitable for building an immune library were used due to the lack of VL domains. Analysis of immune libraries from the genes of the VH3, VHH3 and VH4 families showed that the VHH antibodies share in the VH3 and VH4 gene groups is insignificant, and selection from the combined library is less effective than from the VHH3 family of sequences. We found that the combination of ribosomal and phage displays leads to a higher enrichment of high-affinity fragments and avoids the loss of the original diversity during cloning. The combined method allowed us to obtain a greater number of different high-affinity sequences, and all the tested VHH fragments were able to specifically recognize the target, including the total protein extracts of cell cultures.

scholarly journals Gradient boosting for linear mixed models

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Colin Griesbach ◽  
Benjamin Säfken ◽  
Elisabeth Waldmann
Keyword(s):  
Random Effects ◽  
Mixed Models ◽  
Selection Procedure ◽  
Classification Theory ◽  
Gradient Boosting ◽  
Random Structure ◽  
Boosting Algorithm ◽  
The One ◽  
Biased Estimates ◽  
Selection Of

Abstract Gradient boosting from the field of statistical learning is widely known as a powerful framework for estimation and selection of predictor effects in various regression models by adapting concepts from classification theory. Current boosting approaches also offer methods accounting for random effects and thus enable prediction of mixed models for longitudinal and clustered data. However, these approaches include several flaws resulting in unbalanced effect selection with falsely induced shrinkage and a low convergence rate on the one hand and biased estimates of the random effects on the other hand. We therefore propose a new boosting algorithm which explicitly accounts for the random structure by excluding it from the selection procedure, properly correcting the random effects estimates and in addition providing likelihood-based estimation of the random effects variance structure. The new algorithm offers an organic and unbiased fitting approach, which is shown via simulations and data examples.

scholarly journals Network Analysis Based on Unique Spectral Features Enables an Efficient Selection of Genomically Diverse Operational Isolation Units

2021 ◽  
Vol 9 (2) ◽  
pp. 416
Author(s):  
Charles Dumolin ◽  
Charlotte Peeters ◽  
Evelien De Canck ◽  
Nico Boon ◽  
Peter Vandamme
Keyword(s):  
Network Analysis ◽  
Hierarchical Clustering ◽  
Mass Spectra ◽  
Spectral Features ◽  
Maldi Tof ◽  
Maldi Tof Mass Spectra ◽  
Diversity Studies ◽  
Technical Sample ◽  
Efficient Selection ◽  
Selection Of

Culturomics-based bacterial diversity studies benefit from the implementation of MALDI-TOF MS to remove genomically redundant isolates from isolate collections. We previously introduced SPeDE, a novel tool designed to dereplicate spectral datasets at an infraspecific level into operational isolation units (OIUs) based on unique spectral features. However, biological and technical variation may result in methodology-induced differences in MALDI-TOF mass spectra and hence provoke the detection of genomically redundant OIUs. In the present study, we used three datasets to analyze to which extent hierarchical clustering and network analysis allowed to eliminate redundant OIUs obtained through biological and technical sample variation and to describe the diversity within a set of spectra obtained from 134 unknown soil isolates. Overall, network analysis based on unique spectral features in MALDI-TOF mass spectra enabled a superior selection of genomically diverse OIUs compared to hierarchical clustering analysis and provided a better understanding of the inter-OIU relationships.

Efficient selection of feature sets possessing high coefficients of determination based on incremental determinations

2003 ◽  
Vol 83 (4) ◽  
pp. 695-712 ◽  
Author(s):  
Ronaldo F. Hashimoto ◽  
Edward.R. Dougherty ◽  
Marcel Brun ◽  
Zheng-Zheng Zhou ◽  
Michael L. Bittner ◽  
...  
Keyword(s):  
Feature Sets ◽  
Efficient Selection ◽  
Selection Of
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