High-Throughput Chemical Screening for Inhibitors of Salmonella Island 2

被引:0
作者
Tsai, Caressa N. [1 ,2 ]
Coombes, Brian K. [1 ,2 ]
机构
[1] McMaster Univ, Dept Biochem & Biomed Sci, Hamilton, ON L8S 4L8, Canada
[2] McMaster Univ, Michael G DeGroote Inst Infect Dis Res, Hamilton, ON L8S 4L8, Canada
来源
STAR PROTOCOLS | 2020年 / 1卷 / 02期
基金
加拿大健康研究院;
关键词
EXPRESSION;
D O I
10.1016/j.xpro.2020.100057
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Anti-virulence therapies are under active investigation as antibiotic alternatives; however, their identification from large-scale chemical libraries poses a unique challenge. The dispensability of virulence factors for growth in vitro precludes conventional, optical density-based screening methods. Here, we provide a protocol for high throughput screening with a cell-based, promoter reporter platform. We describe the use of this method for the identification of anti-SPI-2 inhibitors specific to Salmonella Typhimurium, which may be modified to investigate other virulence factors.For complete details on the use and execution of this protocol, please refer to Tsai et al. (2020).
引用
收藏
页数:16
相关论文
共 8 条
[1]   Expression and secretion of Salmonella pathogenicity island-2 virulence genes in response to acidification exhibit differential requirements of a functional type III secretion apparatus and SsaL [J].
Coombes, BK ;
Brown, NF ;
Valdez, Y ;
Brumell, JH ;
Finlay, BB .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (48) :49804-49815
[2]   A robust platform for chemical genomics in bacterial systems [J].
French, Shawn ;
Mangat, Chand ;
Bharat, Amrita ;
Cote, Jean-Philippe ;
Mori, Hirotada ;
Brown, Eric D. .
MOLECULAR BIOLOGY OF THE CELL, 2016, 27 (06) :1015-1025
[3]   Expression of flagella is coincident with uropathogenic Escherichia coli ascension to the upper urinary tract [J].
Lane, M. Chelsea ;
Alteri, Christopher J. ;
Smith, Sara N. ;
Mobley, Harry L. T. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (42) :16669-16674
[4]   A cell-based approach to characterize antimicrobial compounds through kinetic dose response [J].
MacNair, Craig R. ;
Stokes, Jonathan M. ;
French, Shawn ;
Myers, Cullen L. ;
Iyer, Kali R. ;
Brown, Eric D. .
BIOORGANIC & MEDICINAL CHEMISTRY, 2016, 24 (24) :6315-6319
[5]   Rank Ordering Plate Data Facilitates Data Visualization and Normalization in High-Throughput Screening [J].
Mangat, Chand S. ;
Bharat, Amrita ;
Gehrke, Sebastian S. ;
Brown, Eric D. .
JOURNAL OF BIOMOLECULAR SCREENING, 2014, 19 (09) :1314-1320
[6]   Transcriptional Priming of Salmonella Pathogenicity Island-2 Precedes Cellular Invasion [J].
Osborne, Suzanne E. ;
Coombes, Brian K. .
PLOS ONE, 2011, 6 (06)
[7]   Targeting Two-Component Systems Uncovers a Small-Molecule Inhibitor of Salmonella Virulence [J].
Tsai, Caressa N. ;
MacNair, Craig R. ;
Cao, My P. T. ;
Perry, Jordyn N. ;
Magolan, Jakob ;
Brown, Eric D. ;
Coombes, Brian K. .
CELL CHEMICAL BIOLOGY, 2020, 27 (07) :793-+
[8]   A simple statistical parameter for use in evaluation and validation of high throughput screening assays [J].
Zhang, JH ;
Chung, TDY ;
Oldenburg, KR .
JOURNAL OF BIOMOLECULAR SCREENING, 1999, 4 (02) :67-73