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KMID : 0525719990040030137
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Journal of Chitin and Chitosan
1999 Volume.4 No. 3 p.137 ~ p.143
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Isolation and Identification of Chitinase cDNA from Entomopathogenic Fungus , Metarhizium anisopliae
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이동규/Lee DG
김철호/Kim CH
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Abstract
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A chitinase cDNA was isolated from the entomopathogenic fungus Metarhizium anisopliae which is a natural pathogen to many insect pests. The chitinase is one of the cuticle-degrading enzyme and was presumably recognized as a virulence factor in insect mycosis. The CDNA library was constructed with RNAs isolated from M. anisopliae mycellia cultured in medium containing chitin as a sole carbon and nitrogen source. The chitinase clone was directly screened from cDNA library grown on NZY agar media containing glycol chitin [0.01%(w/v)] by staining with Calcofluor white M2R. The chitinase gene fused with the lacZ gene was functionally expressed in Escherichia coli. The molecular weight of the fusion protein was about 70 kDa judged on 10% SDS-Polyacrylamide gel. The chitinase gene appears to be a single copy in the M. αnisopliae genome as the result of genomic Southern analysis.
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