弓形虫章鱼碱脱氢酶Tg ODH多克隆抗体的制备与初步应用OA

Toxoplasma gondii,an important zoonotic parasitic protozoan belonging to the apicomplexans,exhibits remarkable plas-ticity in its central carbon metabolism,thereby enabling its adaptation to diverse parasitic environments.Although several studies have elucidated the important roles of metabolic enzymes in Toxoplasma proliferation and dissemination,the functions of a subset of these enzymes remain uncharacterized.Herein,we focused on the T.gondii octopine dehydrogenase protein(ODH).Bioinformatic analysis revealed that ODH is a stable hydrophilic protein lacking signal peptides and transmembrane domains,and containing two conserved functional domains:an octopine dehydrogenase domain(208-378 aa)and a prephenate dehydrogenase domain(78-123 aa).We gen-erated polyclonal antibodies in mice immunized with a specific short peptide.Western blotting detected a specific band around 50 kDa,which was consistent with the predicted molecular weight of ODH.Indirect immunofluorescence assays further demonstrated the pro-tein's localization predominantly to the parasite cytoplasm.Together,these results indicated the successful production of mouse-derived polyclonal antibodies against ODH and delineated its subcellular localization,thereby providing a foundation for further investi-gation of the biological roles of ODH in T.gondii.