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HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS
oleh: ANTONIUS SUWANTO, AMARILA MALIK, BUDI TJAHJONO, ROB HARLING
| Format: | Article |
|---|---|
| Diterbitkan: | SEAMEO, BIOTROP 2011-11-01 |
Deskripsi
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