HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS

Authors

  • ANTONIUS SUWANTO Department of Biology, Faculty of Mathematics and Sciences,
  • AMARILA MALIK Southeast Asian Regional Center for Tropical Biology (SEAMEO BIOTROP), Bogor,
  • BUDI TJAHJONO Department of Plant Pests and Diseases, Faculty of Agriculture, Bogor Agricultural University, Bogor-16144, Indonesia: '
  • ROB HARLING Department of Crop Science. Scottish Agricultural College, Edinburgh EH9 3JG, UK.

DOI:

https://doi.org/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

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