Department of Pathogen Biology, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Public Health, Southern Medical University, Guangzhou, 510515, China.
Experimental Animal Center, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China.
Parasit Vectors. 2019 Mar 15;12(1):112. doi: 10.1186/s13071-019-3378-y.
Aromatic amino acid hydroxylase 2 (AAH2) is a bradyzoite-specific upregulated protein that may alter host behaviour by altering the host dopaminergic pathway. To better understand the role of the parasite's AAH2 in host-parasite interactions, we generated an AAH2 fluorescent marker strain of T. gondii using the TALEN technique.
We generated an AAH2 fluorescent marker strain of T. gondii, which was designated PRU/AAH2-eGFP, using the TALEN technique. This strain stably expressed pyrimethamine resistance for screening and expressed enhanced green fluorescent protein (eGFP)-tagged AAH2 in the bradyzoite stage. The bradyzoite conversion of PRU/AAH2-eGFP was observed both in vitro and in vivo. The fluorescence localization of AAH2 in mouse models of chronic infection was observed by a Bruker in vivo imaging system.
Transgenic T. gondii was successfully generated by the TALEN system. The eGFP-tagged AAH2 could be detected by in vivo imaging.
This study verified the feasibility of using TALEN technology for T. gondii research and provided an in vivo imaging method for in vivo research of bradyzoite-stage proteins.
芳香族氨基酸羟化酶 2（AAH2）是一种速殖子特异性上调的蛋白，它可能通过改变宿主的多巴胺能途径来改变宿主的行为。为了更好地了解寄生虫的 AAH2 在宿主-寄生虫相互作用中的作用，我们使用 TALEN 技术生成了 T. gondii 的 AAH2 荧光标记株。
我们使用 TALEN 技术生成了 T. gondii 的 AAH2 荧光标记株，命名为 PRU/AAH2-eGFP。该菌株稳定表达了氨苯砜抗性，用于筛选，并在速殖子阶段表达了增强型绿色荧光蛋白（eGFP）标记的 AAH2。在体外和体内观察到 PRU/AAH2-eGFP 的速殖子转化。使用 Bruker 体内成像系统观察到慢性感染小鼠模型中 AAH2 的荧光定位。
通过 TALEN 系统成功生成了转基因 T. gondii。可以通过体内成像检测到 eGFP 标记的 AAH2。
本研究验证了使用 TALEN 技术进行 T. gondii 研究的可行性，并提供了一种用于速殖子期蛋白体内研究的体内成像方法。
