title: Cloning of the gene for glutamate decarboxylase and its expression during conidiation in Neurospora crassa.
abstract: Neurospora crassa glutamate decarboxylase (GAD) is produced during conidiation and stored in dormant conidia. Polyclonal antibody was generated to GAD that had been purified to homogeneity. The anti-GAD antibody was specific for N. crassa GAD and inhibited GAD activity. The level of GAD protein decreased during conidial germination, indicating that GAD was degraded during this phase of development. The anti-GAD antibody was used to isolate a cDNA clone of GAD from a lambda ZAP cDNA expression library. Escherichia coli containing a plasmid with the cDNA insert produced GAD activity. The cDNA clone contained a 2.6 kbp insert and hybridized to a 2.6 kb mRNA species from conidiating cultures of N. crassa. GAD mRNA was not present in vegetative hyphae. In conidiating cultures, GAD mRNA was first detected when conidia began to appear. The level of GAD mRNA increased as conidiation progressed. This is the first example of the cloning of an enzyme that is regulated at the level of mRNA during the asexual developmental cycle of N. crassa.
authors.affiliation: Southern Illinois University, Carbondale 62901.
authors.forename: R
authors.initials: R
authors.lastname: Hao
authors.forename: J C
authors.initials: JC
authors.lastname: Schmit
mesh_terms: D000906:Antibodies; D003001:Cloning, Molecular; D004247:DNA; D004926:Escherichia coli; D015870:Gene Expression; D005968:Glutamate Decarboxylase / Q000235:genetics* / Q000276:immunology; D009492:Neurospora crassa / Q000201:enzymology* / Q000254:growth & development; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D000906:Antibodies; D012333:RNA, Messenger; D004247:DNA; D005968:Glutamate Decarboxylase