biostudies-literatureSugi TNCRR NIH HHSMinistry of Education, Culture, Sports, Science and TechnologyNational Institutes of HealthJapan Science and Technology AgencyBio-oriented Technology Research Advancement InstitutionJapan Society for the Promotion of Science93-101https://www.ebi.ac.uk/biostudies/studies/S-EPMC3862444biostudies-literatureUnknown3Toxoplasma gondii is an important food and waterborne pathogen that causes severe disease in immunocompromised patients. Bumped kinase inhibitors (BKIs) have an antiparasitic effect on T. gondii tachyzoite growth by targeting T. gondii calmodulin-domain protein kinase 1 (TgCDPK1). To identify mutations that confer resistance to BKIs, chemical mutagenesis was performed, followed by selection in media containing either 250 or 1000 nM 1NM-PP1. Whole-genome sequence analysis of resistant clones revealed single nucleotide mutations in T. gondii mitogen-activated protein kinase 1 (TgMAPK1) at amino acids 162 (L162Q) and 171 (I171N). Plasmid constructs having the TgMAPK1 L162Q mutant sequence successfully replaced native TgMAPK1 genome locus in the presence of 1000 nM 1NM-PP1. The inhibitory effeInternational journal for parasitology. Drugs and drug resistanceIdentification of mutations in TgMAPK1 of Toxoplasma gondii conferring resistance to 1NM-PP1.PMC3862444P41 RR-01081P41 RR001081Murakoshi FHorimoto TKato KAkashi HRecuenco FCTakemae HSugi TIwanaga TKobayashi KGong HIshiwa AfalseIdentification of mutations in TgMAPK1 of Toxoplasma gondii conferring resistance to 1NM-PP1.Toxoplasma gondii is an important food and waterborne pathogen that causes severe disease in immunocompromised patients. Bumped kinase inhibitors (BKIs) have an antiparasitic effect on T. gondii tachyzoite growth by targeting T. gondii calmodulin-domain protein kinase 1 (TgCDPK1). To identify mutations that confer resistance to BKIs, chemical mutagenesis was performed, followed by selection in media containing either 250 or 1000 nM 1NM-PP1. Whole-genome sequence analysis of resistant clones revealed single nucleotide mutations in T. gondii mitogen-activated protein kinase 1 (TgMAPK1) at amino acids 162 (L162Q) and 171 (I171N). Plasmid constructs having the TgMAPK1 L162Q mutant sequence successfully replaced native TgMAPK1 genome locus in the presence of 1000 nM 1NM-PP1. The inhibitory effe2013-01-01T00:00:00Z2013 Dec2026-05-04T05:24:55.709Z2019-03-27T01:18:27ZS-EPMC38624442453329810.1016/j.ijpddr.2013.04.001