Isolation of recombinant MVA using F13L selection
Modified vaccinia virus Ankara (MVA) has become a widely used vector for vaccine and laboratory purposes. Despite significant advances in recombinant MVA technology, the isolation of recombinant viruses remains a tedious and difficult process. This chapter describes the use of an efficient and easy-...
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Published in | Methods in molecular biology (Clifton, N.J.) Vol. 890; p. 93 |
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Format | Journal Article |
Language | English |
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United States
2012
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Abstract | Modified vaccinia virus Ankara (MVA) has become a widely used vector for vaccine and laboratory purposes. Despite significant advances in recombinant MVA technology, the isolation of recombinant viruses remains a tedious and difficult process. This chapter describes the use of an efficient and easy-to-use selection system adapted for MVA. The system is based on the requirement of the viral gene F13L for efficient virus spread in cell culture, which results in a severe block in virus transmission when F13L gene is deleted (Blasco R, Moss B. J Virol 65:5910-5920, 1991; Blasco R, Moss B. J Virol 66:4170-4179, 1992). The insertion of foreign genes in the MVA genome is accomplished by recombination of a transfected plasmid carrying the foreign genes and the F13L with the genome of an F13L knockout virus. Subsequently, selection of virus recombinants is carried out by serial passage and/or plaque purification of viruses that have recovered the F13L gene. |
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AbstractList | Modified vaccinia virus Ankara (MVA) has become a widely used vector for vaccine and laboratory purposes. Despite significant advances in recombinant MVA technology, the isolation of recombinant viruses remains a tedious and difficult process. This chapter describes the use of an efficient and easy-to-use selection system adapted for MVA. The system is based on the requirement of the viral gene F13L for efficient virus spread in cell culture, which results in a severe block in virus transmission when F13L gene is deleted (Blasco R, Moss B. J Virol 65:5910-5920, 1991; Blasco R, Moss B. J Virol 66:4170-4179, 1992). The insertion of foreign genes in the MVA genome is accomplished by recombination of a transfected plasmid carrying the foreign genes and the F13L with the genome of an F13L knockout virus. Subsequently, selection of virus recombinants is carried out by serial passage and/or plaque purification of viruses that have recovered the F13L gene. |
Author | Sánchez-Puig, Juana M Lorenzo, María M Blasco, Rafael |
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Snippet | Modified vaccinia virus Ankara (MVA) has become a widely used vector for vaccine and laboratory purposes. Despite significant advances in recombinant MVA... |
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SubjectTerms | Animals Base Sequence Cells, Cultured Cloning, Molecular Cricetinae DNA, Recombinant Gene Knockout Techniques Genetic Markers Membrane Proteins - genetics Promoter Regions, Genetic Transduction, Genetic Vaccinia virus - genetics Vaccinia virus - growth & development Vaccinia virus - isolation & purification Viral Envelope Proteins - genetics Viral Load Virus Cultivation |
Title | Isolation of recombinant MVA using F13L selection |
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