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HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS

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Author(s): AMARILA MAHK | ANTONIUS SUWANTO | BUDI TJAHJONO | BUDI TJAHJONO

Journal: BIOTROPIA : the Southeast Asian Journal of Tropical Biology
ISSN 0215-6334

Volume: 20;
Start page: 36;
Date: 2003;
Original page

Keywords: Aeromonas caviae/ chitinase gene/transcriptional fusion/PKm'V Vtac-chilPseudomonas fluorescens

ABSTRACT
A transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR). The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli carrying the recombinant plasmids with Pseudomotws fluorescens 5100, a phyllosphere bacterium, was performed. Pseudomonas fluorescens 5100 exconjugants were examined for constitutive expression of chitinase employing a spectrophotometric assay; they showed stronger chitin degradation activity than Escherichia coli transformants. Using a fungal antagonism plate assay, this chitinolytic P. fluorescens, however, could not inhibit selected phytopathogenic fungi.
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