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Gene Therapy

Neurospora-expression-system

HIV protease inhibitors useful for the treatment of AIDS

OF THE INVENTION AND PREFERRED EMBODIMENTS This invention is concerned with compounds of Formula I,...

Continuously operational biological reactor

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Making a lightly oxidized starch additive by adding a cationic polymer to starch slurry prior to heating the slurry

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Glucaric acid monoamides and their use to prepare poly(glucaramides)

OF THE INVENTION The novel alkali metal N-aminohydrocarbyl- and N-aminoheterohydrocarbyl-D-...

Process for preparing poly (glucaramides)

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High efficiency retroviral vector which contains genetically engineered cellular non-coding sequence harboring splicing acceptor

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Nucleic acid probes and amplification oligonucleotides for Neisseria species

OF THE INVENTION A. Definitions The following terms have the indicated meanings in the ...

Site-specific protein modification

OF THE INVENTION Site specific modification of proteins and peptides provides a powerful research ...

Neurospora expression system

Details

Inventors: Stuart, W. Dorsey; Ivy, John M.; Koo, Kenneth;
Assignee: Hawaii Biotechnology Group, Inc. (Aiea, HI)
Primary Examiner: Ketter; James
Assistant Examiner:
Attorney, Agent or Firm: Rae-Venter Law Group, P.C.

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.

DETAILED DESCRIPTION What is claimed is: 1.
A method of producing a mammalian peptide in a Neurospora fungus host, said method comprising: transforming by DNA uptake Neurospora spheroplasts with (1) a DNA construct comprising as operably linked components a constitutive promoter functional in said fungus host, a first DNA sequence encoding a mammalian peptide under the transcriptional regulation of said promoter and a transcriptional termination regulatory region functional in said host, and (2) a second DNA sequence comprising a marker gene which is expressed in said host; selecting for transformed host cells comprising said construct by means of said marker gene; identifying stably transformed host cells; and growing said stably transformed host cells whereby said first DNA sequence is expressed, and said cells produce said mammalian peptide.
2.
A method of producing a mammalian peptide in a Neurospora crassa host, said method comprising: transforming by DNA uptake N.
crassa spheroplasts with (1) a DNA construct comprising as operably linked components a .
beta.
-tubulin promoter functional in said host, a first DNA sequence encoding a mammalian peptide under the transcriptional regulation of said .
beta.
-tubulin promoter and a transcriptional termination regulatory region functional in said host, and (2) a second DNA sequence comprising a marker gene which is expressed in said host; selecting for transformed host cells comprising said construct by means of said marker gene; identifying stably transformed host cells; and growing said stably transformed host cells whereby said first DNA sequence is expressed, and said cells produce said mammalian peptide.
3.
A DNA construct comprising a Neurospora .
beta.
-tubulin promoter and a DNA sequence encoding a mammalian peptide under the transcriptional control of said promoter.
4.
A DNA construct according to claim 3, wherein said mammalian peptide is preprochymosin.
5.
A stably transformed Neurospora cell containing a DNA construct comprising as operably linked components

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