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Cloning and expression of a chitinase gene from the hyperparasitic fungus Aphanocladium album.
Institut de Génétique et Microbiologie, Université Paris Sud, Orsay, France.
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.
PMID: 1735126 [PubMed - indexed for MEDLINE]
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Cited by 3 PubMed Central articles
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Expression of two major chitinase genes of Trichoderma atroviride (T. harzianum P1) is triggered by different regulatory signals.
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Appl Environ Microbiol. 1999 May; 65(5):1858-63.
[Appl Environ Microbiol. 1999]
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Isolation of genomic DNAs from plants, fungi and bacteria using benzyl chloride.
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Nucleic Acids Res. 1993 Nov 11; 21(22):5279-80.
[Nucleic Acids Res. 1993]
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A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies, and infected plant tissues.
Möller EM, Bahnweg G, Sandermann H, Geiger HH.
Nucleic Acids Res. 1992 Nov 25; 20(22):6115-6.
[Nucleic Acids Res. 1992]