From GFP to β-lactamase: advancing intact cell imaging for toxins and effectors. Pathog Dis 2015 Dec;73(9):ftv097
Date
10/27/2015Pubmed ID
26500183Pubmed Central ID
PMC4732026DOI
10.1093/femspd/ftv097Scopus ID
2-s2.0-85021388978 (requires institutional sign-in at Scopus site) 7 CitationsAbstract
Canonical reporters such as green fluorescent protein (GFP) and luciferase have assisted researchers in probing cellular pathways and processes. Prior research in pathogenesis depended on sensitivity of biochemical and biophysical techniques to identify effectors and elucidate entry mechanisms. Recently, the β-lactamase (βlac) reporter system has advanced toxin and effector reporting by permitting measurement of βlac delivery into the cytosol or host βlac expression in intact cells. βlac measurement in cells was facilitated by the development of the fluorogenic substrate, CCF2-AM, to identify novel effectors, target cells, and domains involved in bacterial pathogenesis. The assay is also adaptable for high-throughput screening of small molecule inhibitors against toxins, providing information on mechanism and potential therapeutic agents. The versatility and limitations of the βlac reporter system as applied to toxins and effectors are discussed in this review.
Author List
Zuverink M, Barbieri JTAuthor
Joseph T. Barbieri PhD Professor in the Microbiology and Immunology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Cytological TechniquesFluorescence
Genes, Reporter
Green Fluorescent Proteins
Humans
Optical Imaging
beta-Lactamases
