摘要
利用CRISPR/Cas9技术成功构建了弓形虫Ⅱ型Pru虫株的TGME49_228400定位株和缺失株(PruΔ228400),研究了TGME49_228400在弓形虫中的亚细胞定位和基本生物学功能。Western-blot结果显示TGME49_228400蛋白大小为69.9 ku。间接免疫荧光检测显示,TGME49_228400定位于顶质体,TGME49_228400缺失后对弓形虫顶质体的形态无影响。与Pru野生株相比,TGME49_228400缺失株形成噬斑和增殖的能力均显著减弱(P<0.01),而入侵、逸出、体外缓殖子转化能力和对小鼠的毒力无显著差异(P>0.05)。本研究结果可为后续阐明弓形虫中其他含WD40结构域蛋白的功能和作用机制奠定基础。
A localization strain and PruΔ228400 strain of Toxoplasma gondii typeⅡPru strain were successfully constructed using the CRISPR/Cas9 technology and the subcellular localization and basic biological function of TGME49_228400 in T.gondii were analyzed.Western-blot showed that the size of TGME49_228400 protein was 69.9 ku.Immunofluorescence analysis results revealed that TGME49_228400localized in the apicoplast.Deletion of TGME49_228400 had no effect on morphology of the apicoplast.Compared to the wild type Pru strain,the deletion of TGME49_228400 exhibited a significant reduction of the ability to form plaques and replication(P<0.01),but no effect on the invasion,egress,the in vitro bradyzoite differentiation and virulence in mice of T.gondii(P<0.05).The results may provide a basis for the subsequent elucidation of the functions and mechanisms of action of other WD40-containing domain proteins in T.gondii.
作者
张天宇
孙黎秀
祁淑芸
郑晓楠
张芝玮
王金磊
王萌
袁安文
ZHANG Tian-yu;SUN Li-xiu;QI Shu-yun;ZHENG Xiao-nan;ZHANG Zhi-wei;WANG Jin-lei;WANG Meng;YUAN An-wen(College of Veterinary Medicine,Hunan Agricultural University,Changsha 410000,China;State Key Laboratory for Animal Disease Control and Prevention/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2023年第10期1298-1306,共9页
Chinese Veterinary Science
基金
国家自然科学基金青年项目(32002306)。
