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US 7,067,251 B2Granted
Methods of directly selecting cells expressing inserts of interest
AssigneeUniversity of Rochester
InventorsMaurice Zauderer, Ernest S. Smith
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Abstract
The present invention relates to a high efficiency method of introducing DNA into linear DNA viruses such as poxvirus, a method of producing libraries in linear DNA viruses such as poxvirus, and methods of selecting polynucleotides of interest based on cell nonviability or other phenotypes.
Claims51
A method of selecting a target polynucleotide, comprising:
- (a) introducing into a population of mammalian host cells a library of insert polynucleotides;
- wherein at least one of said insert polynucleotides comprises the target polynucleotide;
- wherein said library is constructed in a vaccinia virus vector using trimolecular recombination and wherein expression of said target polynucleotide directly or indirectly promotes host cell death;
- (b) culturing said host cells under conditions such that said insert polynucleotides are expressed;
- and
- (c) collecting insert polynucleotides from those host cells which undergo cell death;
- wherein said cell death is not the result of a cytotoxic T lymphocyte-induced lytic event.
4 more elements
Dependencies are inferred from the claim text.