弓形虫TGME49_240850基因编码的含WD40结构域蛋白的生物学功能OA北大核心

In order to explore the biological function of the WD40-containing domain protein encoded by the TGME49_240850 gene from Toxoplasma gondii,a conditional knockdown strain(TGME49_240850-mAID-6HA)was successfully constructed based on the CRISPR/Cas9 technology.The subcellular localization and biological function of TGME49_240850 from T.gondii were analyzed.Immunofluorescence analysis revealed that the TGME49_240850 protein was localized in the nucleus,and was successfully degraded upon adding indole-3-acetic acid(IAA).Western-blot showed that the size of TGME49_240850-mAID-6HA protein was approximately 90 kDa.Compared to the control group,the TGME49_240850-mAID-6HA strain showed a signifi-cant reduction in the ability to form plaques(P＜0.001)and replication(P＜0.01)under the IAA condi-tion,but no effect on the invasion,egress,and in vitro bradyzoite differentiation of T.gondii(P＞0.05).These results proved that the degradation of TGME49_240850 reduced the ability of replication in T.gondii.In conclusion:a conditional knockdown strain for TGME49_240850 was successfully constructed,which laid a foundation for further research on the function of WD40-containing domain-related proteins in T.gondii.