ROB HARLING
Department of Crop Science. Scottish Agricultural College, Edinburgh EH9 3JG, UK.

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HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS ANTONIUS SUWANTO; AMARILA MALIK; BUDI TJAHJONO; ROB HARLING
BIOTROPIA - The Southeast Asian Journal of Tropical Biology No. 20 (2003)
Publisher : SEAMEO BIOTROP

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (405.968 KB) | DOI: 10.11598/btb.2003.0.20.182

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