Molecular cloning of a Neurospora crassa carotenoid biosynthetic gene (albino-3) regulated by blue light and the products of the white collar genes.
The albino-3 (al-3) gene of Neurospora crassa, which probably encodes the carotenoid biosynthetic enzyme geranylgeranyl pyrophosphate synthetase, was cloned. The N. crassa triple mutant al-3 qa-2 aro-9 was transformed to qa-2+ with mixtures of plasmids bearing N. crassa DNA inserts, and the transformants were screened for the al-3+ phenotype. One al-3+ qa-2+ transformant (AL3-1) was examined in detail and shown to contain intact vector sequences integrated into the N. crassa genome. The vector and some flanking sequences were recovered from AL3-1 after restriction, ligation, and selection of chloramphenicol-resistant transformants of Escherichia coli. The flanking sequences were subsequently used to detect the al-3-containing plasmid in the mixture of about 1,800 plasmids. Restriction fragment length polymorphism mapping was carried out to confirm the identity of the cloned fragment. The level of the al-3 mRNA was shown to be increased 15-fold in light-induced (compared with that in dark-grown) wild-type mycelia. The light-dependent increase in al-3 mRNA levels was not observed in presumed regulatory mutant (white collar) strains.
Nelson
M A
MA
Dipartimento Biopatologia Umana, Universita di Roma La Sapienza, Policlinico Umberto I, Rome, Italy.
Morelli
G
G
Carattoli
A
A
Romano
N
N
Macino
G
G
D002338:Carotenoids / Q000096:biosynthesis / Q000235:genetics*; D003001:Cloning, Molecular; D004122:Dimethylallyltranstransferase / Q000235:genetics; D005786:Gene Expression Regulation / Q000528:radiation effects; D005800:Genes, Fungal; D005822:Genetic Vectors; D008027:Light; D009154:Mutation; D009491:Neurospora / Q000235:genetics*; D009492:Neurospora crassa / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D010858:Pigmentation / Q000235:genetics; D010957:Plasmids; D012331:RNA, Fungal / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015183:Restriction Mapping
D012331:RNA, Fungal; D012333:RNA, Messenger; D002338:Carotenoids; D004122:Dimethylallyltranstransferase