| Literature DB >> 24991689 |
Tatsuki Sugi1, Tatsunori Masatani2, Fumi Murakoshi3, Shin-ichiro Kawazu1, Kentaro Kato4.
Abstract
Toxoplasma gondii can differentiate into tachyzoites or bradyzoites. To accelerate the investigation of bradyzoite differentiation mechanisms, we constructed a reporter parasite, PLK/DLUC_1C9, for a high-throughput assay. PLK/DLUC_1C9 expressed firefly luciferase under the bradyzoite-specific BAG1 promoter. Firefly luciferase activity was detected with a minimum of 10(2) parasites induced by pH 8.1. To normalize bradyzoite differentiation, PLK/DLUC_1C9 expressed Renilla luciferase under the parasite's α-tubulin promoter. Renilla luciferase activity was detected with at least 10(2) parasites. By using PLK/DLUC_1C9 with this 96-well format screening system, we found that the protein kinase inhibitor analogs, bumped kinase inhibitors 1NM-PP1, 3MB-PP1, and 3BrB-PP1, had bradyzoite-inducing effects.Entities:
Keywords: Bradyzoite; DUAL luciferase assay; High-throughput screening; Reporter parasite; Toxoplasma gondii
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Year: 2014 PMID： 24991689 DOI： 10.1016/j.ab.2014.06.018
Source DB: PubMed Journal: Anal Biochem ISSN： 0003-2697 Impact factor: 3.365