Transient transfection of Cryptosporidium parvum using green fluorescent protein (GFP) as a marker

W. Li; N. Zhang; X. Liang; J. Li; P. Gong; X. Yu; G. Ma; U.M. Ryan; X. Zhang

doi:10.1016/j.molbiopara.2009.07.003

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Transient transfection of Cryptosporidium parvum using green fluorescent protein (GFP) as a marker

Journal article

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Transient transfection of Cryptosporidium parvum using green fluorescent protein (GFP) as a marker

W. Li, N. Zhang, X. Liang, J. Li, P. Gong, X. Yu, G. Ma, U.M. Ryan and X. Zhang

Molecular and Biochemical Parasitology, Vol.168(2), pp.143-148

2009

DOI: https://doi.org/10.1016/j.molbiopara.2009.07.003

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Abstract

Cryptosporidium parvum is a protozoan parasite that infects a variety of mammals. The parasite has been shown to harbor a dsRNA virus (CPV) and in the present study, we have developed a CPV transient transfection system for this parasite by using green fluorescent protein (GFP) to replace the partial gene encoding region of the larger dsRNA (CPV-L) and the smaller dsRNA (CPV-S) virus. Two viral RNA-mediated transfection vectors: pCPVL-GFP and pCPVS-GFP were successfully constructed and both in vitro transcripts were electroporated into oocysts and sporozoites. Transient expression of GFP was detected in C. parvum oocysts and excysted sporozoites by fluorescence microscopy and by RT-PCR detection of GFP mRNA and antisense RNA in transfected C. parvum oocysts. Our study provides a new approach for studying gene expression and regulation in C. parvum and will hopefully lead to the construction of a stable CPV transfection system in the future.

Details

Title: Transient transfection of Cryptosporidium parvum using green fluorescent protein (GFP) as a marker
Authors/Creators: W. Li (Author/Creator) - Jilin University
N. Zhang (Author/Creator) - Jilin University
X. Liang (Author/Creator) - Jilin University
J. Li (Author/Creator) - Jilin University
P. Gong (Author/Creator) - Jilin University
X. Yu (Author/Creator) - Jilin University
G. Ma (Author/Creator) - China Rural Technology Development Center
U.M. Ryan (Author/Creator) - Murdoch University
X. Zhang (Author/Creator) - Jilin University
Publication Details: Molecular and Biochemical Parasitology, Vol.168(2), pp.143-148
Publisher: Elsevier B.V.
Identifiers: 991005542625407891
Copyright: © 2009 Elsevier B.V.
Murdoch Affiliation: School of Veterinary and Biomedical Sciences
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.246 Diarrheal Diseases; 1.246.985 Cryptosporidium
Web Of Science research areas: Biochemistry & Molecular Biology; Parasitology
ESI research areas: Microbiology