Construction Of DNA Probe
DNA fragments specific to the cDNA of the ferritin of Acacia mangium were amplified by the polymerase chain reaction (PCR). The two primers were as described below:
The cDNA for the template was synthesized with an RT-PCR Kit (Stratagene). The amplified DNA fragment was subcloned into pT7 Blue Vector (Stratagene) and used as a probe to screen for the cDNA of ferritin.
DIG DNA labeling and Detection System Kit was used to screen for ferittin cDNA. Plaques were screened by plaque hybridization according to kit instructions. After a second screening, the inserted DNA fragment of positive plaques was subcloned into pUC118 and sequenced by a PRISM Dye Terminator Cycle Sequencing Kit (Perkin Elmer) with an automated DNA sequencer (model 373; Perkin Elmer).
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