(1,3) .beta.-glucan synthase genes and inducible inhibition of fungal growth using the antisense constructs derived therefrom
OwnerClaude P. Selitrennikoff, Carol S. Enderlininventors
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Abstract
A (1,3).beta.-glucan synthase mutant useful in the research and development of anti-fungal drugs. A key step in fungal wall assembly, hyphal growth and infection is the synthesis of (1,3).beta.-linked glucan. Because .beta.-glucan is not made by humans, (1,3).beta.-glucan synthase is an attractive target for development of new antifungal antibiotics. A (1,3).beta.-glucan synthase gene has been cloned and the cDNA sequence and polypeptide amino acid sequence of the gene has been established.
Claims16
Purified DNA encoding (1,3).beta.-glucan synthase wherein said (1,3).beta.-glucan synthase has an amino acid sequence of SEQ ID NO:2.
Purified DNA comprising a fragment of the sequence of SEQ ID NO:
- 1 and encoding a protein having (1,3).beta.-glucan svnthase catalytic activity.
Purified DNA encoding (1.3).beta.-glucan synthase and having the sequence of SEQ ID NO:1.
A process for cloning a (1,3).beta.-glucan synthase gene comprising the steps of:
- (a) generating (1,3).beta.-glucan synthase mutants from a temperature-sensitive osmotic mutant of Neurospora crassa;
- (b) cultivating cell-wall-less (1,3).beta.-glucan synthase mutants from said (1,3).beta.-glucan synthase mutants;
- (c) preparing pools of cDNA from a Neurospora crassa genomic library, said cDNA being located on a cosmid which has a hygromycin-resistance gene;
- (d) transforming the (1,3).beta.-glucan synthase mutants with the cDNA to form transformants;
- (e) identifying the transformants by selecting for the hygromycin-resistance gene on said cosmid;
- and
- (f) isolating an individual cosmid that complements said (1,3).beta.-glucan synthase mutants using a sib selection procedure to yield a cloned (1,3).beta.-glucan synthase gene.
Dependencies are inferred from the claim text.