Rhoptry protein 5(ROP5) Is a Key Virulence Factor inNeospora caninum.
Neospora caninum, of theApicomplexaphylum, is a common cause of abortions in cattle and nervous system dysfunction in dogs. Rhoptry proteins ofApicomplexaplay an important role in virulence. The objectives of this study were to study functions of NcROP5 inN. caninumby deleting the NcROP5 gene from the wild Nc-1 strain. We selectedNcROP5in ToxoDB and successfully constructed anNcROP5gene-deleted vector, pTCR-NcROP5-CD KO. Then we screened the NcROP5 knockout strains (ΔNcROP5) at the gene, protein and transcription levels. Plaque assay, host cell invasion assay and intracellular proliferation test showed that the ΔNcROP5 strain had less plaque space, weakened invasion capacity and slower intracellular growth. Animal testing showed significantly lower cerebral load of ΔNcROP5 than the load of the Nc-1 strain, as well as a loss of virulence for the ΔNcROP5 strains. Phenotypic analyses using the label-free LC-MS/MS assay-based proteomic method and KEGG pathway enrichment analysis showed a reduction ofNcGRA7transcription and altered expression of multiple proteins including the apicomplexan family of binding proteins. The present study indicated that ROP5 is a key virulence factor inN. caninumin mice. The proteomic profiling of Nc-1 and ΔNcROP5 provided some data on differential proteins. These data provide a foundation for future research of protein functions inN. caninum.
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University Beijing, China.
Lei
L
Ma
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University Beijing, China.
Jing
J
Liu
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University Beijing, China.
Muzi
M
Li
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University Beijing, China.
Yong
Y
Fu
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University Beijing, China.
Xiao
X
Zhang
Key Laboratory of Animal Epidemiology and Zoonosis, Ministry of Agriculture, National Animal Protozoa Laboratory, College of Veterinary Medicine, China Agricultural University Beijing, China.
Qun
Q
Liu
Neospora caninum; gene editing; protozoan parasites; rhoptry protein 5; virulence factor