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Published on: November 1, 2011
Reciprocal complementation of bovine parainfluenza virus type 3 lacking either the membrane or fusion gene
Marina Takada1, Ryosuke Matsuura1, Takehiro Kokuho2
1Laboratory of Environmental Microbiology, Faculty of Medicine, University of Tsukuba, Ibaraki 305-8575, Japan.
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.
Insights
Researchers developed two defective bovine parainfluenza virus type 3 (BPIV3) strains for efficient protein expression. This reciprocal complementation method enables robust viral vector applications in cell cultures.
Area of Science:
- Virology
- Molecular Biology
- Biotechnology
Background:
- Bovine parainfluenza virus type 3 (BPIV3) is a significant pathogen.
- Developing efficient viral vectors is crucial for biotechnology and research.
- Existing BPIV3 vectors have limitations in expressing large or multiple proteins.
Purpose of the Study:
- To create and characterize two defective BPIV3 strains for reciprocal complementation.
- To establish a novel viral vector system for enhanced protein expression in cell cultures.
- To demonstrate the utility of this system for expressing large or multiple proteins.
Main Methods:
- Generation of two BPIV3 strains: one lacking the M gene (ΔM-EGFP) and another lacking the F gene (ΔF-mSB), with fluorescent protein expression.
- Trans-complementation of defective viral proteins in Madin-Darby bovine kidney (MDBK) cells.
- Co-infection of MDBK cells with both defective strains at low multiplicity of infection (MOI).
Main Results:
- Easily obtained complemented viruses upon co-infection with ΔM-EGFP and ΔF-mSB.
- Complemented viruses exhibited growth efficiency comparable to wild-type BPIV3.
- Successful viral passaging in MDBK cells at MOI as low as 0.01.
- Evidence suggests the formation of multiploid virus particles containing both defective genomes.
Conclusions:
- The reciprocal complementation of defective BPIV3 strains is an effective method for generating functional viral vectors.
- This approach facilitates the expression of large or multiple proteins in cell cultures.
- The developed BPIV3 vector system offers a promising tool for paramyxovirus-based applications.
