Abstract
BackgroundAromatic 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.MethodsWe 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.ResultsTransgenic T. gondii was successfully generated by the TALEN system. The eGFP-tagged AAH2 could be detected by in vivo imaging.ConclusionsThis 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.
Highlights
Aromatic amino acid hydroxylase 2 (AAH2) is a bradyzoite-specific upregulated protein that may alter host behaviour by altering the host dopaminergic pathway
The SAG1 gene promoter was amplified by PCR and inserted into pTALEN-L digested with AscI/SpeI
The GRA2 gene terminator was amplified via PCR and inserted into pTALEN-L-SAG1 digested with BglII/NotI
Summary
Aromatic amino acid hydroxylase 2 (AAH2) is a bradyzoite-specific upregulated protein that may alter host behaviour by altering the host dopaminergic pathway. Toxoplasma gondii is an obligate intracellular protozoan parasite and one of the most widespread zoonotic parasites, and it can infect most warm-blooded animals and humans [1, 2]. This species infects up to a third of the world’s population [3, 4] and represents a serious threat to public health. Most healthy adults usually show symptoms of latent infection after infection with T. gondii At this time, the parasites convert to bradyzoites and exist in the form of cysts in the brain and muscle tissue of the host [5], which causes latent infection and may lead to changes in several functions of the host brain and behaviour [6].
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