Cloning and expression of a chitinase gene from the hyperparasitic fungus Aphanocladium album.
Recombinant clones from a cDNA library of an Aphanocladium album chitinase-overproducing mutant strain were isolated by screening with antiserum against a 39 kDa chitinase purified from this hyperparasitic fungus. Analysis of the isolated positive clones indicated that most of them carried the same cDNA. A cDNA from this group was used as a hybridization probe to isolate an 8 kb DNA fragment from a genomic library of the wild-type strain. The chitinase 1 gene was mapped to this fragment by two independent approaches. Its partial DNA sequence was in perfect agreement with an amino-terminal peptide sequence obtained by sequencing 23 amino acids of the 39 kDa chitinase. Its transfer in Fusarium oxysporum resulted in a transformant producing both a protein of about 39 kDa that cross-reacted with the chitinase antiserum and a chitinase activity that was inhibited by the same antiserum. Northern blot analysis indicates that the cloned chitinase gene was subject to catabolite repression and appeared inducible by chitin.
Blaiseau
P L
PL
Institut de Génétique et Microbiologie, Université Paris Sud, Orsay, France.
Kunz
C
C
Grison
R
R
Bertheau
Y
Y
Brygoo
Y
Y
D000595:Amino Acid Sequence; D001483:Base Sequence; D002686:Chitin / Q000494:pharmacology; D002688:Chitinases / Q000235:genetics*; D003001:Cloning, Molecular; D005670:Fusarium / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D015723:Gene Library; D005800:Genes, Fungal; D005947:Glucose / Q000494:pharmacology; D003904:Mitosporic Fungi / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D014170:Transformation, Genetic
D002686:Chitin; D002688:Chitinases; D005947:Glucose