中国兽医科学2025，Vol.55Issue(5)：577-585,9.DOI:10.16656/j.issn.1673-4696.2025.0079
弓形虫TGME49_240850基因编码的含WD40结构域蛋白的生物学功能
Biological function of WD40-containing domain protein encoded by TGME49_240850 gene from Toxoplasma gondii
摘要
Abstract
In order to explore the biological function of the WD40-containing domain protein encoded by the TGME49_240850 gene from Toxoplasma gondii,a conditional knockdown strain(TGME49_240850-mAID-6HA)was successfully constructed based on the CRISPR/Cas9 technology.The subcellular localization and biological function of TGME49_240850 from T.gondii were analyzed.Immunofluorescence analysis revealed that the TGME49_240850 protein was localized in the nucleus,and was successfully degraded upon adding indole-3-acetic acid(IAA).Western-blot showed that the size of TGME49_240850-mAID-6HA protein was approximately 90 kDa.Compared to the control group,the TGME49_240850-mAID-6HA strain showed a signifi-cant reduction in the ability to form plaques(P＜0.001)and replication(P＜0.01)under the IAA condi-tion,but no effect on the invasion,egress,and in vitro bradyzoite differentiation of T.gondii(P＞0.05).These results proved that the degradation of TGME49_240850 reduced the ability of replication in T.gondii.In conclusion:a conditional knockdown strain for TGME49_240850 was successfully constructed,which laid a foundation for further research on the function of WD40-containing domain-related proteins in T.gondii.关键词
弓形虫/WD40结构域/条件性敲除/生长素诱导降解系统Key words
Toxoplasma gondii/WD40 domain/conditional knockdown/auxin-inducible degradation system分类
农业科技引用本文复制引用
洪照微,王萌,孙黎秀,寇永杰,马志雅,江新成,付宝权..弓形虫TGME49_240850基因编码的含WD40结构域蛋白的生物学功能[J].中国兽医科学,2025,55(5):577-585,9.基金项目
国家重点研发计划项目(2023YFD1801000) （2023YFD1801000）
甘肃省自然科学基金项目(23JRRA553) （23JRRA553）
西藏自治区科技计划项目(QYXTZX-RKZ2024-03-3) （QYXTZX-RKZ2024-03-3）
