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| Cloning of peroxidase gene in taro |
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| DOI：10.16968/j.issn.1004-894X.2016.09.006 |
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| Abstract: |
| The taro peroxidase gene sequence was amplified using degenerate primers which were designed
by iCODEHOP according to the taro codon usage frequency. And the peroxidase gene non-flanking sequence was
amplified successfully. Then the peroxidase gene flanking sequences were amplified by hiTAIL-PCR primers which
were designed on the basis of the non-flanking sequence. As a result，1 165 bp sequences were successfully cloned
from taro genomic DNA，and it was affirmed that they were part of the sequences of peroxidase gene by homology
analysis. Then by hiTAIL-PCR technology，the peroxidase gene flanking sequences were amplified，and 689 bp
sequences were amplified. The Sequencing result could be spliced together with the 1 165 bp sequences. And the
amplified taro peroxidase gene sequences spliced together with its flanking sequences were 1 854 bp. Sequence
alignment with known peroxidase，we found that there were amino acid coding sequences on both sides of the
amplification sequences. So，the hiTAIL-PCR technology was used as the second time，and 165 bp sequences were
amplified. Spliced together with the 1 854 bp sequences，the amplified taro peroxidase gene sequences were 2 019
bp. With the gene on softberry prediction software analysis，it was found that the sequences contained 4 peroxidase
exons. Homology analysis showed that the peroxidase gene was Class III heme-containing peroxidase. |
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