Methods and compositions for combinatorial-based discovery of new multimeric molecules
AssigneeUniversity of Hawaii
InventorW. Dorsey Stuart
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Abstract
The present invention prowdes methods and compositions for producing and screening combinatorial libraries of multimeric proteins. The present invention relies on the use of filamentous fungal heterokaryons that are produced using two or more parent strains into which a population of DNA molecules encoding variants of a multimeric protein have been introduced.
Claims13
A panel comprising two or more heterokaryons wherein each of said heterokaryons produces a different variant of a multimeric protein and each heterokaryon is formed by fusing a first and a second fungal parent strain, wherein said heterokaryon requires the presence of both fungal parent nuclei for survival, said first and said second parent fungal strain each contain an exogenously supplied nucleic acid molecule that encodes a variant of a subunit of a multimeric protein and where said first and said second parent strains are homozygous for all heterokaryon compatibility alleles.
A method for producing a panel of heterokaryons comprising two or more heterokaryons wherein each of said heterokaryons produces a multimeric protein comprising the steps of:
- fusing two or more pairs of fungal strains, each pair consisting of a first and second fungal parent strain, said first and said second parent fungal strains containing an exogenously supplied nucleic acid molecule that encodes a variant of a subunit of a multimeric protein and where said first and said second parent strains are homozygous for all heterokaryon compatibility alleles and
- selecting two or more heterokaryons thus formed to generate said panel,
- wherein said heterokaryon requires the presence of both fungal parent nuclei for survival.
Dependencies are inferred from the claim text.