| Autor: |
Sánchez-Puig JM; Departamento de Biotecnología, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain., Lorenzo MM, Blasco R |
| Jazyk: |
angličtina |
| Zdroj: |
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2012; Vol. 890, pp. 93-111. |
| DOI: |
10.1007/978-1-61779-876-4_5 |
| Abstrakt: |
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. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
|
