Evaluation of a Yeast Two-Hybrid Library by High-Throughput Sequencing

2020 ◽  
Vol 19 (8) ◽  
pp. 3567-3572
Author(s):  
Qi Yu ◽  
Yueheng Hu ◽  
Jiangshuo Su ◽  
Peiling Li ◽  
Luyao Zhang ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Sequencing ◽  
Yeast Two Hybrid ◽  
Two Hybrid

scholarly journals Unraveling protein interactions between the temperate virus Bam35 and its Bacillus host using an integrative yeast two hybrid-high throughput sequencing approach

2021 ◽  
Author(s):  
Ana Lechuga ◽  
Cédric Lood ◽  
Mónica Berjón-Otero ◽  
Alicia Del Prado ◽  
Jeroen Wagemans ◽  
...  
Keyword(s):  
High Throughput ◽  
Protein Interactions ◽  
High Throughput Sequencing ◽  
Genomic Library ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Protein Protein Interaction ◽  
Novel Approach ◽  
Diverse Groups ◽  
Two Hybrid

Bacillus virus Bam35 is the model Betatectivirus and member of the Tectiviridae family, which is composed of tailless, icosahedral, and membrane-containing bacteriophages. The interest in these viruses has greatly increased in recent years as they are thought to be an evolutionary link between diverse groups of prokaryotic and eukaryotic viruses. Additionally, betatectiviruses infect bacteria of the Bacillus cereus group, known for their applications in industry and notorious since it contains many pathogens. Here, we present the first protein-protein interactions network for a tectivirus-host system by studying the Bam35- Bacillus thuringiensis model using a novel approach that integrates the traditional yeast two-hybrid system and Illumina high-throughput sequencing. We generated and thoroughly analyzed a genomic library of Bam35’s host B. thuringiensis HER1410 and screened interactions with all the viral proteins using different combinations of bait-prey couples. In total, this screen resulted in the detection of over 4,000 potential interactions, of which 183 high-confidence interactions were defined as part of the core virus-host interactome. Overall, host metabolism proteins and peptidases are particularly enriched within the detected interactions, distinguishing this host-phage system from the other reported host-phage protein-protein interaction networks (PPIs). Our approach also suggests biological roles for several Bam35 proteins of unknown function, resulting in a better understanding of the Bam35- B. thuringiensis interaction at the molecular level.

scholarly journals Unraveling Protein Interactions between the Temperate Virus Bam35 and Its Bacillus Host Using an Integrative Yeast Two Hybrid–High Throughput Sequencing Approach

2021 ◽  
Vol 22 (20) ◽  
pp. 11105
Author(s):  
Ana Lechuga ◽  
Cédric Lood ◽  
Mónica Berjón-Otero ◽  
Alicia del Prado ◽  
Jeroen Wagemans ◽  
...  
Keyword(s):  
High Throughput ◽  
Protein Interactions ◽  
High Throughput Sequencing ◽  
Genomic Library ◽  
Structural Protein ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Novel Approach ◽  
Host Membrane ◽  
Two Hybrid

Bacillus virus Bam35 is the model Betatectivirus and member of the family Tectiviridae, which is composed of tailless, icosahedral, and membrane-containing bacteriophages. Interest in these viruses has greatly increased in recent years as they are thought to be an evolutionary link between diverse groups of prokaryotic and eukaryotic viruses. Additionally, betatectiviruses infect bacteria of the Bacillus cereus group, which are known for their applications in industry and notorious since it contains many pathogens. Here, we present the first protein–protein interactions (PPIs) network for a tectivirus–host system by studying the Bam35–Bacillus thuringiensis model using a novel approach that integrates the traditional yeast two-hybrid system and high-throughput sequencing (Y2H-HTS). We generated and thoroughly analyzed a genomic library of Bam35′s host B. thuringiensis HER1410 and screened interactions with all the viral proteins using different combinations of bait–prey couples. Initial analysis of the raw data enabled the identification of over 4000 candidate interactions, which were sequentially filtered to produce 182 high-confidence interactions that were defined as part of the core virus–host interactome. Overall, host metabolism proteins and peptidases were particularly enriched within the detected interactions, distinguishing this host–phage system from the other reported host–phage PPIs. Our approach also suggested biological roles for several Bam35 proteins of unknown function, including the membrane structural protein P25, which may be a viral hub with a role in host membrane modification during viral particle morphogenesis. This work resulted in a better understanding of the Bam35–B. thuringiensis interaction at the molecular level and holds great potential for the generalization of the Y2H-HTS approach for other virus–host models.

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

Abstract 3033: Identification of the MUC16/CA125 interaction network: Results of a high-throughput yeast two-hybrid screening

2012 ◽  
Author(s):  
Stephan Polterauer ◽  
Dharmarao Thapi ◽  
Nikolaus Schultz ◽  
Ouathek Ouerfelli ◽  
Nancy Chen ◽  
...  
Keyword(s):  
High Throughput ◽  
Interaction Network ◽  
Yeast Two Hybrid ◽  
Two Hybrid ◽  
Hybrid Screening

Interaction network of antimicrobial peptides of Arabidopsis thaliana, based on high-throughput yeast two-hybrid screening

2012 ◽  
Vol 58 ◽  
pp. 245-252 ◽  
Author(s):  
Coralie Damon ◽  
Julia Dmitrieva ◽  
Yordan Muhovski ◽  
Frédéric Francis ◽  
Laurence Lins ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Antimicrobial Peptides ◽  
High Throughput ◽  
Interaction Network ◽  
Yeast Two Hybrid ◽  
Two Hybrid ◽  
Hybrid Screening

scholarly journals High throughput flow cytometry based yeast two-hybrid array approach for large-scale analysis of protein-protein interactions

2011 ◽  
Vol 81A (1) ◽  
pp. 90-98 ◽  
Author(s):  
Jun Chen ◽  
Mark B. Carter ◽  
Bruce S. Edwards ◽  
Hong Cai ◽  
Larry A. Sklar
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Protein Interactions ◽  
Large Scale ◽  
Scale Analysis ◽  
Protein Protein Interactions ◽  
Yeast Two Hybrid ◽  
Large Scale Analysis ◽  
Two Hybrid

A yEGFP-based reporter system for high-throughput yeast two-hybrid assay by flow cytometry

2008 ◽  
Vol 73A (4) ◽  
pp. 312-320 ◽  
Author(s):  
Jun Chen ◽  
Jianhong Zhou ◽  
Weon Bae ◽  
Claire K. Sanders ◽  
John P. Nolan ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
High Throughput ◽  
Yeast Two Hybrid ◽  
Reporter System ◽  
Two Hybrid

High-Throughput Yeast Two-Hybrid Screening

2011 ◽  
pp. 39-61 ◽  
Author(s):  
George G. Roberts ◽  
Jodi R. Parrish ◽  
Bernardo A. Mangiola ◽  
Russell L. Finley
Keyword(s):  
High Throughput ◽  
Yeast Two Hybrid ◽  
Two Hybrid ◽  
Hybrid Screening
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