Microplate assay for screening Toxoplasma gondii bradyzoite differentiation with DUAL luciferase assay |
| |
| Authors: | Tatsuki Sugi Tatsunori Masatani Fumi Murakoshi Shin-ichiro Kawazu Kentaro Kato |
| |
| Affiliation: | 1. National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan;2. Transboundary Animal Diseases Center, Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima 890-0065, Japan;3. Department of Veterinary Microbiology, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan |
| |
| 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 102 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 102 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. |
| |
| Keywords: | Bradyzoite DUAL luciferase assay High-throughput screening Reporter parasite Toxoplasma gondii |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
