scholarly journals Construction of gateway-compatible yeast two-hybrid vectors for high throughput analysis of protein interaction

2010 ◽  
Vol 9 (9) ◽  
pp. 1392-1396
Author(s):  
Zhu Tingheng ◽  
Wang Weixia ◽  
Wong Hann lin ◽  
Yang Xiao ◽  
Wang Kun ◽  
...  
Keyword(s):  
High Throughput ◽  
Protein Interaction ◽  
Yeast Two Hybrid ◽  
High Throughput Analysis ◽  
Throughput Analysis ◽  
Hybrid Vectors ◽  
Two Hybrid

High-Throughput Yeast Two-Hybrid Assays for Large-Scale Protein Interaction Mapping

Methods ◽  
2001 ◽  
Vol 24 (3) ◽  
pp. 297-306 ◽  
Author(s):  
Albertha J.M. Walhout ◽  
Marc Vidal
Keyword(s):  
High Throughput ◽  
Protein Interaction ◽  
Large Scale ◽  
Yeast Two Hybrid ◽  
Two Hybrid

scholarly journals Yeast Two-Hybrid Systems and Protein Interaction Mapping Projects for Yeast and Worm

Yeast ◽  
2000 ◽  
Vol 1 (2) ◽  
pp. 88-94 ◽  
Author(s):  
Albertha J. M. Walhout ◽  
Simon J. Boulton ◽  
Marc Vidal
Keyword(s):  
Hybrid Systems ◽  
High Throughput ◽  
Protein Interaction ◽  
Protein Interactions ◽  
Large Scale ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Systematic Analysis ◽  
Technical Features ◽  
Two Hybrid

The availability of complete genome sequences necessitates the development of standardized functional assays to analyse the tens of thousands of predicted gene products in high-throughput experimental settings. Such approaches are collectively referred to as ‘functional genomics’. One approach to investigate the properties of a proteome of interest is by systematic analysis of protein–protein interactions. So far, the yeast two-hybrid system is the most commonly used method for large-scale, high-throughput identification of potential protein–protein interactions. Here, we discuss several technical features of variants of the two-hybrid systems in light of data recently obtained from different protein interaction mapping projects for the budding yeastSaccharomyces cerevisiaeand the nematodeCaenorhabditis elegans.

scholarly journals Yeast Two-Hybrid Systems and Protein Interaction Mapping Projects for Yeast and Worm

Yeast ◽  
2000 ◽  
Vol 1 (2) ◽  
pp. 88-94 ◽  
Author(s):  
Albertha J. M. Walhout ◽  
Simon J. Boulton ◽  
Marc Vidal
Keyword(s):  
Hybrid Systems ◽  
High Throughput ◽  
Protein Interaction ◽  
Protein Interactions ◽  
Large Scale ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Systematic Analysis ◽  
Technical Features ◽  
Two Hybrid

The availability of complete genome sequences necessitates the development of standardized functional assays to analyse the tens of thousands of predicted gene products in high-throughput experimental settings. Such approaches are collectively referred to as ‘functional genomics’. One approach to investigate the properties of a proteome of interest is by systematic analysis of protein–protein interactions. So far, the yeast two-hybrid system is the most commonly used method for large-scale, high-throughput identification of potential protein–protein interactions. Here, we discuss several technical features of variants of the two-hybrid systems in light of data recently obtained from different protein interaction mapping projects for the budding yeast Saccharomyces cerevisiae and the nematode Caenorhabditis elegans.

Combination of Free Software, Light Microscopy and Non-specific Dyes for a High-throughput Analysis in Live/Dead Cell Count

2015 ◽  
Vol 11 (4) ◽  
pp. 233-238 ◽  
Author(s):  
Luciano Cardoso ◽  
Suellen Cordeiro ◽  
Marcio Fronza ◽  
Denise Endringer ◽  
Tadeu de Andrade ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Cell Count ◽  
High Throughput ◽  
Free Software ◽  
Dead Cell ◽  
High Throughput Analysis ◽  
Throughput Analysis

scholarly journals Structural Glycoprotein E2 of Classical Swine Fever Virus Interacts with Host Protein Dynactin Subunit 6 (DCTN6) during the Virus Infectious Cycle

2019 ◽  
Vol 94 (1) ◽  
Author(s):  
M. V. Borca ◽  
E. A. Vuono ◽  
E. Ramirez-Medina ◽  
P. Azzinaro ◽  
K. A. Berggren ◽  
...  
Keyword(s):  
Amino Acid ◽  
Protein Interaction ◽  
Hybrid Approach ◽  
Classical Swine Fever ◽  
Host Protein ◽  
Yeast Two Hybrid ◽  
Protein Protein Interaction ◽  
Viral Virulence ◽  
Glycoprotein E2 ◽  
Two Hybrid

ABSTRACT The E2 protein in classical swine fever (CSF) virus (CSFV) is the major virus structural glycoprotein and is an essential component of the viral particle. E2 has been shown to be involved in several functions, including virus adsorption, induction of protective immunity, and virulence in swine. Using the yeast two-hybrid system, we previously identified a swine host protein, dynactin subunit 6 (DCTN6) (a component of the cell dynactin complex), as a specific binding partner for E2. We confirmed the interaction between DCTN6 and E2 proteins in CSFV-infected swine cells by using two additional independent methodologies, i.e., coimmunoprecipitation and proximity ligation assays. E2 residues critical for mediating the protein-protein interaction with DCTN6 were mapped by a reverse yeast two-hybrid approach using a randomly mutated E2 library. A recombinant CSFV mutant, E2ΔDCTN6v, harboring specific substitutions in those critical residues was developed to assess the importance of the E2-DCTN6 protein-protein interaction for virus replication and virulence in swine. CSFV E2ΔDCTN6v showed reduced replication, compared with the parental virus, in an established swine cell line (SK6) and in primary swine macrophage cultures. Remarkably, animals infected with CSFV E2ΔDCTN6v remained clinically normal during the 21-day observation period, which suggests that the ability of CSFV E2 to bind host DCTN6 protein efficiently during infection may play a role in viral virulence. IMPORTANCE Structural glycoprotein E2 is an important component of CSFV due to its involvement in many virus activities, particularly virus-host interactions. Here, we present the description and characterization of the protein-protein interaction between E2 and the swine host protein DCTN6 during virus infection. The E2 amino acid residues mediating the interaction with DCTN6 were also identified. A recombinant CSFV harboring mutations disrupting the E2-DCTN6 interaction was created. The effect of disrupting the E2-DCTN6 protein-protein interaction was studied using reverse genetics. It was shown that the same amino acid substitutions that abrogated the E2-DCTN6 interaction in vitro constituted a critical factor in viral virulence in the natural host, domestic swine. This highlights the potential importance of the E2-DCTN6 protein-protein interaction in CSFV virulence and provides possible mechanisms of virus attenuation for the development of improved CSF vaccines.

Multiwell Combinatorial Hydrogel Array for High-Throughput Analysis of Cell–ECM Interactions

2021 ◽  
Author(s):  
Ruoxing Lei ◽  
Erin A. Akins ◽  
Kelly C. Y. Wong ◽  
Nicole A. Repina ◽  
Kayla J. Wolf ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Analysis ◽  
Throughput Analysis

Development of pattern recognition based on the nanosheet-DNA probes and an extendible DNA library

The Analyst ◽  
2021 ◽  
Author(s):  
Jiawei Qi ◽  
Pinhua Rao ◽  
Lele Wang ◽  
Li Xu ◽  
Yanli Wen ◽  
...  
Keyword(s):  
Pattern Recognition ◽  
High Throughput ◽  
Analysis Data ◽  
Dna Probes ◽  
High Throughput Analysis ◽  
Throughput Analysis ◽  
Dna Library ◽  
Array Sensing

Pattern recognition, also called “array sensing” is a recognition strategy with a wide and expandable analysis range, based on the high-throughput analysis data. In this work, we constructed a sensor...

Highly Sensitive Detection of Paraquat with Pillar[5]arene as an aptamer in α-Hemolysin Nanopore

2021 ◽  
Author(s):  
Xiaojia Jiang ◽  
Mingsong Zang ◽  
Fei Li ◽  
Chunxi Hou ◽  
Quan Luo ◽  
...  
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Single Molecule ◽  
Single Molecule Detection ◽  
Sensitive Detection ◽  
High Throughput Analysis ◽  
Throughput Analysis ◽  
The Real ◽  
Highly Sensitive ◽  
Highly Sensitive Detection

Biological nanopore-based techniques have attracted more and more attention recently in the field of single-molecule detection, because they allow the real-time, sensitive, high-throughput analysis. Herein, we report an engineered biological...

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