heterologous expression of a chitinase gene from aeromonas caviaein pseudomonas fluorescens

heterologous expression of a chitinase gene from aeromonas caviaein pseudomonas fluorescens

;ANTONIUS SUWANTO;AMARILA MAHK;BUDI TJAHJONO;BUDI TJAHJONO
journal of advanced veterinary research 2003 Vol. 20 pp. 36-48
210
suwanto2003biotropia:heterologous

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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