Vol 49 No. 4: 361-366 [PDF] [Full Text]
Identification and characterization of GRA6/GRA7 of Neospora caninum in MDBK cells
Jingquan Dong, Jianhua Li, Jinpeng Wang, Fei Li, Ju Yang, Pengtao Gong*, He Li, and Xichen Zhang*
College of Veterinary Medicine, Jilin University, Changchun 130062, China
These authors contributed equally to this work.

Abstract  Neospora caninum, an apicomplexan parasite, is recognized as a major bovine abortifacient. Dense granule antigens (GRAs) play important roles in the formation and modification of parasitophorous vacuoles (PVs) in Toxoplasma gondii. However, a few studies investigating GRAs have been reported in N. caninum. The aim of the present study was to characterize the dense GRA6/GRA7 of N. caninum in PVs using MDBK cells as a host cell model. Neospora caninum was inoculated into MDBK cells, and changes were observed using a transmission electron microscope (TEM). Neospora caninum GRA6/GRA7 were identified and characterized using bioinformatics, cell fractionation, and immunofluorescence. The TEM results revealed that integrated PVs were present in MDBK cells after N. caninum infection. Bioinformatics analysis showed that NcGRA6/NcGRA7 shared 28.76% and 29.66% homology with T. gondii GRA6/GRA7 (TgGRA6/TgGRA7) but had similar signal peptides, transmembrane domains, and motifs. Cell fractionation and subcellular localization analyses both showed that NcGRA6 was distributed in the lumen and intravacuolar network in soluble and transmembrane forms. The transmembrane form of NcGRA7 was observed in the PV membrane. These data lay a foundation for further study on bovine neosporosis and NcGRA6/NcGRA7 function during PV formation.


Keywords   Neospora caninum, dense granule antigen, MDBK cells, subcellular localization


Received   2016-11-7  


Funding  This work was supported by the grants from the National Basic Science Research Program of China (973 Program) (No. 2015CB150300) and the Special Fund for Agro-scientific Research in the Public Interest in China (No. 20130304).


* Correspondence address  Tel/Fax: +86-431-87981351; E-mail: gongpt@jlu.edu.cn (P.G.)/xczhang@jlu.edu.cn (X.Z.)

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