Plasmids and method for obtaining viral particles
AssigneeUniversidad de Santiago de Chile
InventorsCarolina Beltrán Pavez, Marcelo Cortez San Martin, Eugenio Spencer Ossa, Carolina Tambley Zamorano, Daniela Toro Ascuy
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Abstract
The present invention describes a viral RNA expression plasmid and a method for obtaining viral particles based on the plasmids comprising transfecting animal cells with an expression vector or a set of expression vectors capable of expressing a nucleoprotein and RNA-dependent polymerase RNA; and transfecting the animal cell with an expression vector or a set of expression vectors with nucleotide sequences encoding recombinant RNA molecules.
Claims13
A method of obtaining viral particles, the method comprising:
- a) transfecting each of a plurality of fish cells with a set of plasmids, which express:
- SEQ ID NO 7, SEQ ID NO 8, SEQ ID NO 9, and SEQ ID NO 10, wherein one of SEQ ID NOs 7, 8, 9, or 10 is cloned individually in each of the plasmids;
- b) once the cells are transfected in step a), co-transfecting the fish cells of step a) with a set of eight plasmids:
- pSS-URG/1, pSS-URG/2, pSS-URG/3, pSS-URG/4, pSS-URG/5, pSS-URG/6, pSS-URG/7, and pSS-URG/8, each plasmid having:
- a promoter sequence of Salmo salar internal transcribed sequences (ITS-1) region;
- a hammerhead ribozyme (HH rib) sequence;
- one coding sequence of infectious salmon anemia virus (ISAV) inserted in a cloning site sequence, wherein each pSS-URG/1 to 8 plasmid includes a different coding sequence of ISAV;
- a hepatitis δ virus ribozyme sequence;
- and
- a rabbit β-globin transcription terminator sequence;
- and
- c) obtaining recombinant viral particles resulting from steps a) and b).
A set of expression plasmids including SEQ ID NOs 7, 8, and 9, wherein each of SEQ ID NOs 7, 8, and 9 is cloned individually in each expression plasmid.
An expression plasmid including SEQ ID NO 10.
Dependencies are inferred from the claim text.