University of Hawai'i

Hawaii Biotechnology Group, Inc.

W Dorsey Stuart

Mark B Frank

John M Ivy

Kenneth Koo

Elie K Kato

The present invention provides improved host cells for use in producing secreted recombinant proteins, methods of making the improved hosts and uses thereof. The invention specifically provides strains of Neurospora crassa which produce reduced levels of extracellular proteases when compared with wild-type Neurospora crassa.

Neurospora hosts for the production of recombinant proteins, and methods

The al-3 and related promoters can be used to provide light-regulated recombinant production of heterologous proteins in filamentous fungi. Expression systems utilizing these promoters can be placed in vectors which also optionally contain selectable marker means.

Light-regulated promoters for production of heterologous proteins in

Neurospora crassa transformations are employed for production of heterologous proteins, where DNA constructs are prepared using a foreign gene under transcriptional and translational regulatory regions functional in Neurospora. A functional signal sequence is provided for secretion of the heterologous gene product. An auxotrophic Neurospora host is employed and transformed by the combination of the subject construct and a complementing gene. The resulting transformants provide for efficient secretion of the heterologous product.

Neurospora expression system

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.

Methods and compositions for combinatorial-based discovery of new

The invention relates to a heterokaryotic filamentous fungus host capable of producing a heterologous heterodimer comprising at least two subunits. The heterokaryon contains a first and second nucleus; each nucleus contains an expression system for one subunit of the heterodimer. The heterokaryon is prepared by culturing together a first fungus host strain and a second fungus host strain that is homozygous with the first fungus host strain with respect to all heterokaryon compatibility alleles, wherein the first and second fungus host strains are cultured together under conditions wherein neither the first nor the second fungus strain can survive unless the heterokaryotic host is formed.

Heterologous dimeric proteins produced in heterokaryons

Monoclonal antibodies are provided capable of distinguishing DNA-RNA hybrid complexes from single stranded DNA and RNA and double stranded DNA and RNA. The antibodies find particular use in determining the presence of a specific nucleic acid sequence on a solid surface. Single stranded polynucleotide is fixed to a solid (gel) surface and then hybridized with the complementary probe. The hybrid complex specific monoclonal antibody is then added to bind to any hybrid complexes which have formed. By appropriate label, the hybrid complex may be visualized in a variety of ways.

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Kaneohe, HI, United States of America

W Dorsey Stuart