Rescue of recombinant peste des petits ruminants virus: creation of a GFP-expressing virus and application in rapid virus neutralization test.
Peste des petits ruminants virus (PPRV) causes high mortality in goats and sheep and the disease has shown a greatly increased geographic distribution over the last 15 years. It is responsible for serious socioeconomic problems in some of the poorest developing countries. The ability to create recombinant PPRV would provide a useful tool for investigating the biology of the virus and the pathology of disease, as well as for developing new vaccines and diagnostic methods. Here we report the first successful rescue of recombinant PPRV from a full-length cDNA clone of the virus genome. Successful recovery of PPRV was achieved by using a RNA polymerase II promoter to drive transcription of the full-length virus antigenome. We have used this technique to construct a virus expressing a tracer protein (green fluorescent protein, GFP). The recombinant virus replicated as well as the parental virus and could stably express GFP during at least 10 passages. The newly established reverse genetics system for PPRV provides a novel method for constructing a vaccine using PPRV as a vector, and will also prove valuable for fundamental research on the biology of the virus. We found that our recombinant virus allowed more rapid and higher throughput assessment of PPRV neutralization antibody titer via the virus neutralization test (VNT) compared with the traditional method.
College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China. chenweiye1980@yahoo.com.cn.
Qianqian
Q
Hu
Weiye
W
Chen
Kehe
K
Huang
Michael D
MD
Baron
Zhigao
Z
Bu
D000818:Animals; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018076:DNA, Complementary; D016679:Genome, Viral; D015511:Goat Diseases / Q000276:immunology* / Q000821:virology; D006041:Goats; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009500:Neutralization Tests / Q000662:veterinary; D016366:Open Reading Frames; D029021:Peste-des-Petits-Ruminants / Q000517:prevention & control / Q000662:veterinary*; D018107:Peste-des-petits-ruminants virus / Q000235:genetics* / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary; D012756:Sheep; D012757:Sheep Diseases / Q000276:immunology / Q000821:virology; D014162:Transfection / Q000662:veterinary; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology; D014709:Vero Cells; D014765:Viral Vaccines / Q000235:genetics / Q000276:immunology*
D018076:DNA, Complementary; D014614:Vaccines, Synthetic; D014765:Viral Vaccines; D049452:Green Fluorescent Proteins