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唐金凤，鲁耀邦*，桂柳姿.mRNA 差异显示法筛选黄花石蒜中加兰他敏合成相关基因[J].湖南中医药大学学报,2012,32(7):34-36[点击复制] |
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| mRNA 差异显示法筛选黄花石蒜中加兰他敏合成相关基因 |
| 唐金凤，鲁耀邦*，桂柳姿 |
| (湖南中医药大学药学院，中药药性与药效三级科研实验室，省中药现代化研究实验室) |
| 摘要: |
| 目的筛选和克隆黄花石蒜加兰他敏相关基因。方法采用mRNA 差异显示法（mRNA differential display
PCR，DD-PCR）， 对7 月份黄花石蒜产加兰他敏含量最高的部位花蕊和含量最低的部位花葶进行基因表达差异的研
究，筛选其差异片段，再对差异片段进行回收、克隆、测序及序列分析和同源性比较。结果黄花石蒜花蕊和花葶中存
在明显的基因表达差异，发现差异条带44 条，经序列分析和同源性比较，确认其中2 个条带与黄花石蒜加兰他敏合成
相关，另一条可能与加兰他敏运输代谢调控有关。结论通过DD-PCR 得到的3 个同源序列为研究黄花石蒜中加兰他
敏的合成途径提供了有力依据。 |
| 关键词: 黄花石蒜 加兰他敏 mRNA 差异显示技术 生物信息学 |
| DOI： |
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| 基金项目:湖南省自然科学基金资助项目(10JJ2024)；湖南省教育厅科研基金资助项目(07A052)。 |
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| Screening of the gene related to galantamine biosynthesis of Lycoris aurea Herb. by mRNA differential display |
| TANG Jin-feng， LU Yao-bang， GUI Lui-zi |
| (TCM Uniwersity of Hunan) |
| Abstract: |
| Objective To isolate and clone the gene related to galantamine biosynthesis of Lycoris
aurea Herb. Methods The specially expression cDNA fragments of the gene related to galantamine
biosynthesis were screened by differential display polymerase chain reaction(DD-PCR) after extracting
the total RNA from the pistils tissues and the scape tissues of the Lycoris aurea
Herb cropped in July which had differential contents of the galantamine. Then these fragments
were separately cloned to plasmid PMD19 -T and subsequently underwent sequence analysis
and homogenous comparison. Results 44 differential display bands were found in gene expression
between the pistils tissues and the scape tissues of the Lycoris aurea Herb. According to the sequence
analysis and homologenous comparison， two of them were confirmed to related
with galantamine biosynthesis， one of them was possible related with galantamine transport regulation.
Conclusion The three homologous sequences of Lycoris aurea screened by DD-PCR provide
strong basis of galantamine synthetic pathways. |
| Key words: Lycoris aurea Herb galantamine mRNA DD-PCR bioinformatics |
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