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J Virol Methods. 2017 Nov;249:25-30. doi: 10.1016/j.jviromet.2017.08.010. Epub 2017 Aug 24.

Reciprocal complementation of bovine parainfluenza virus type 3 lacking either the membrane or fusion gene.

Author information

1
Laboratory of Environmental Microbiology, Faculty of Medicine, University of Tsukuba, Ibaraki 305-8575, Japan.
2
Biologics Production, Center for Animal Disease Control and Prevention, National Institute of Animal Health, NARO, Tsukuba, Ibaraki 305-0856, Japan.
3
Viral Diseases and Epidemiology Research Division, National Institute of Animal Health, NARO, Tsukuba, Ibaraki 305-0856, Japan.
4
Laboratory of Environmental Microbiology, Faculty of Medicine, University of Tsukuba, Ibaraki 305-8575, Japan. Electronic address: ktakeuch@md.tsukuba.ac.jp.

Abstract

Two defective bovine parainfluenza virus type 3 (BPIV3) strains were generated, one lacking the membrane (M) protein gene and expressing EGFP (ΔM-EGFP) and the other lacking the fusion (F) protein gene and expressing mStrawberry (ΔF-mSB), by supplying deficient proteins in trans. When Madin-Darby bovine kidney (MDBK) cells were co-infected with ΔM-EGFP and ΔF-mSB at a multiplicity of infection (MOI) of 0.1, complemented viruses were easily obtained. Complemented viruses grew as efficiently as wild-type BPIV3 and could be passaged in MDBK cell cultures even at an MOI of 0.01, possibly due to multiploid virus particles containing genomes of both ΔM-EGFP and ΔF-mSB. This reciprocal complementation method using two defective viruses would be useful to express large or multiple proteins in cell cultures using paramyxovirus vectors.

KEYWORDS:

BPIV3; Complementation; Defective virus

PMID:
28842134
DOI:
10.1016/j.jviromet.2017.08.010
[Indexed for MEDLINE]

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