Keyword search (3,619 papers available)


Using Models to (Re-)Design Synthetic Circuits.

Author(s): McCallum G, Potvin-Trottier L

Mathematical models play an important role in the design of synthetic gene circuits, by guiding the choice of biological components and their assembly into novel gene networks. Here, we present a guide for biologists to build and utilize models of gene netw...

Article GUID: 33405217

Computer-Aided Design of Active Pseudoknotted Hammerhead Ribozymes.

Author(s): Najeh S, Zandi K, Djerroud S, Kharma N, Perreault J

Methods Mol Biol. 2021;2167:91-111 Authors: Najeh S, Zandi K, Djerroud S, Kharma N, Perreault J

Article GUID: 32712917

Metabolomic and lipidomic analyses of chronologically aging yeast.

Author(s): Richard VR, Bourque SD, Titorenko VI

Methods Mol Biol. 2014;1205:359-73 Authors: Richard VR, Bourque SD, Titorenko VI

Article GUID: 25213255

A Cell-Free Content Mixing Assay for SNARE-Mediated Multivesicular Body-Vacuole Membrane Fusion.

Author(s): Karim MA, Samyn DR, Brett CL

Methods Mol Biol. 2019;1860:289-301 Authors: Karim MA, Samyn DR, Brett CL

Article GUID: 30317513

Visualization of SNARE-Mediated Organelle Membrane Hemifusion by Electron Microscopy.

Author(s): Mattie S, Kazmirchuk T, Mui J, Vali H, Brett CL

Methods Mol Biol. 2019;1860:361-377 Authors: Mattie S, Kazmirchuk T, Mui J, Vali H, Brett CL

Article GUID: 30317518

Identification of Genes Involved in the Degradation of Lignocellulose Using Comparative Transcriptomics.

Author(s): Gruninger RJ, Reid I, Forster RJ, Tsang A, McAllister TA

Methods Mol Biol. 2017;1588:279-298 Authors: Gruninger RJ, Reid I, Forster RJ, Tsang A, McAllister TA

Article GUID: 28417376

Isolation and Preparation of Extracellular Proteins from Lignocellulose Degrading Fungi for Comparative Proteomic Studies Using Mass Spectrometry

Author(s): Robert J Gruninger

Fungi utilize a unique mechanism of nutrient acquisition involving extracellular digestion. To understand the biology of these microbes, it is important to identify and characterize the function of proteins that are secreted and involved in this process. Ma...

Article GUID: 28417377

Introduction: Overview of Fungal Genomics.

Author(s): de Vries RP, Grigoriev IV, Tsang A

Methods Mol Biol. 2018;1775:1-7 Authors: de Vries RP, Grigoriev IV, Tsang A

Article GUID: 29876804

Fungal Genomic DNA Extraction Methods for Rapid Genotyping and Genome Sequencing.

Author(s): Bellemare A, John T, Marqueteau S

Methods Mol Biol. 2018;1775:11-20 Authors: Bellemare A, John T, Marqueteau S

Article GUID: 29876805

Mass Spectrometry-Based Proteomics

Author(s): Marcos Rafael Di Falco

Proteomics is the large-scale analysis of proteins rendered possible by modern mass spectrometry analysis methods capable of identifying thousands of peptides/proteins in a fast high-throughput manner. Here I describe protocols for the preparation of fungal...

Article GUID: 29876812

Evolutionary Adaptation to Generate Mutants.

Author(s): de Vries RP, Lubbers R, Patyshakuliyeva A, Wiebenga A, Benoit-Gelber I

Methods Mol Biol. 2018;1775:133-137 Authors: de Vries RP, Lubbers R, Patyshakuliyeva A, Wiebenga A, Benoit-Gelber I

Article GUID: 29876815

Manual Gene Curation and Functional Annotation.

Author(s): McDonnell E, Strasser K, Tsang A

Methods Mol Biol. 2018;1775:185-208 Authors: McDonnell E, Strasser K, Tsang A

Article GUID: 29876819

Evaluating Programs for Predicting Genes and Transcripts with RNA-Seq Support in Fungal Genomes.

Author(s): Reid I

Methods Mol Biol. 2018;1775:209-227 Authors: Reid I

Article GUID: 29876820

Phylogenetic Analysis of Protein Family.

Author(s): Song L, Wu S, Tsang A

Phylogenetic Analysis of Protein Family.

Methods Mol Biol. 2018;1775:267-275

Authors: Song L, Wu S, Tsang A

Abstract
With the number of sequenced genomes increasing rapidly, it is impractical to perform functional and stru...

Article GUID: 29876824


Title:A Cell-Free Content Mixing Assay for SNARE-Mediated Multivesicular Body-Vacuole Membrane Fusion.
Authors:Karim MASamyn DRBrett CL
Link:https://www.ncbi.nlm.nih.gov/pubmed/30317513?dopt=Abstract
DOI:10.1007/978-1-4939-8760-3_19
Category:Methods Mol Biol
PMID:30317513
Dept Affiliation: BIOLOGY
1 Department of Biology, Concordia University, Montréal, QC, Canada.
2 Department of Cell Biology, University of Alberta, Edmonton, AB, Canada.
3 Department of Biology, Concordia University, Montréal, QC, Canada. christopher.brett@concordia.ca.

Description:

A Cell-Free Content Mixing Assay for SNARE-Mediated Multivesicular Body-Vacuole Membrane Fusion.

Methods Mol Biol. 2019;1860:289-301

Authors: Karim MA, Samyn DR, Brett CL

Abstract

Endocytosis is a fundamental process underlying diverse eukaryotic physiology. The terminal stage of this process is membrane fusion between the perimeter membrane of a late endosome filled with intraluminal vesicles, or multivesicular body (MVB), and the lysosome membrane to facilitate catabolism of internalized biomaterials or surface polytopic proteins. To comprehensively understand the mechanisms underlying MVB-lysosome membrane fusion, we developed a quantitative, cell-free assay to study this SNARE-mediated event in molecular detail using Saccharomyces cerevisiae and its vacuolar lysosome, or vacuole, as models. This involves separately isolating organelles from two yeast strains each expressing a different complementary fusion probe targeted to the lumen of either MVBs or vacuoles. Isolated organelles are mixed in vitro under fusogenic conditions. Upon MVB-vacuole membrane fusion, luminal contents mix to facilitate probe interaction, reconstituting ß-lactamase activity recorded by a colorimetric enzyme activity assay. This method accommodates a multitude of approaches (e.g., genetics, addition of purified protein reagents) to study this process in isolation, and in theory could be repurposed to study other SNARE-mediated fusion events within cells.

PMID: 30317513 [PubMed - indexed for MEDLINE]