弓形虫含MFS结构域蛋白TGME49_226020的基本生物学功能研究OA

In order to explore the basic biological function of the major facilitator superfamily(MFS)domain-containing protein encoded by Toxoplasma gondii TGME49_226020 gene,CRISPR-Cas9 technology combined with minimum auxin-inducible degron system was used to construct conditional knockout strains on T.gondii RH-TIR1 and Pru-TIR1 strains,respectively.Indirect immunofluorescence assay showed that TGME49_226020 protein was localized in cytoplasm and was successfully degraded after adding indole acetic acid(IAA).The ability of plaque-forming,invasion,intracellular replication was significantly decreased by successful degradation of TGME49_226020,but without impact on egress ability.The organelle stability test showed that the degradation of TGME49_226020 had no significant effect on the organelle stability of T.gondii.The bradyzoite transformation ability of RH-TIR1-TGME49_226020-mAID strain and Pru-TIR1-TGME49_226020-mAID strain was significantly reduced compared with the corresponding RH-TIR1 wild strain and Pru-TIR1 wild strain after adding IAA.Conclusion:considering that TGME49_226020 is a transport protein containing MFS domain,the degradation of TGME49_226020 protein may affect the normal operation of several metabolic transport pathways in T.gondii,and the underlying mechanism warrants further studies.