| 莫俊杰，梁钾贤，胡汉桥，黄 红，陈 妤.芋头过氧化物酶基因克隆[J].广东农业科学,2016,43(9):37-43 |
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芋头过氧化物酶基因克隆 |
| Cloning of peroxidase gene in taro |
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| DOI：10.16968/j.issn.1004-894X.2016.09.006 |
| 中文关键词: 芋头 过氧化物酶基因 hiTAIL-PCR 克隆 |
| 英文关键词: taro peroxidase gene hiTAIL-PCR cloning |
| 基金项目:国家公益性行业（农业）科研专项经费子课题分项目（200903017-08） |
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| 摘要点击次数: 1860 |
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| 中文摘要: |
| 根据芋头的密码子使用频率，用 iCODEHOP 设计简并引物扩增芋头过氧化物酶基因序列，待
芋头过氧化物酶基因非侧翼序列扩增成功后，依据其序列合成 hiTAIL-PCR 引物扩增芋头过氧化物酶基因
的两侧序列。结果从芋头基因组DNA 中克隆出 1 165 bp 序列，经同源性分析为过氧化物酶基因的部分序
列；再利用hiTAIL-PCR 技术可以扩增出芋头过氧化物酶基因的两侧序列，扩增序列为 689 bp，测序结果
能与1 165 bp 序列拼接到一起，长1 854 bp。与已知的过氧化物酶进行序列比对，发现两侧还有氨基酸的编
码序列，因此进行第2 次hiTAIL-PCR，扩增出165 bp 芋头过氧化物酶基因的侧翼序列，拼接后得到2 019
bp。用Softberry 上的基因预测软件分析后，发现包含过氧化物酶4 个外显子，同源性分析表明，此过氧化物
酶基因为含血红素的第三类过氧化物酶。 |
| 英文摘要: |
| 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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