| 黄湘文1,2 张冲2 石新国2 陈华2 郑奕雄1,2.花生RGA 片段的克隆及初步分析[J].,2011,11(9):1631-1633 |
| 花生RGA 片段的克隆及初步分析 |
| Cloning and Primary Analysis for RGA Fragments in Peanut |
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| DOI： |
| 中文关键词: 花生 RGA 同源克隆 |
| 英文关键词: Peanut RGA homology cloning |
| 基金项目:国家863 计划花生抗病基因的克隆与功能研究（2006AA10A115） |
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| 中文摘要: |
| 目的：通过同源克隆获得了花生闽花6 号的RGA 片段，为其抗性的研究及抗性育种提供了参考资料。方法：试验分为两组：
其一通过利用抗性基因的NBS 保守区所设计的简并引物对花生品种闽花六号进行了RGA 片段扩增，其二结合已登录的花生
RGA 片段序列经过多元比对后设计简并引物进行RGA 片段的扩增及序列分析；分析比较两组克隆方法的效果。结果：测序分析
表明：前者20 条随机测序序列中没有一条与已知RGA 片段序列相似；后者20 条随机测序序列中有18 条为RGA 片段序列，其
登录号为GenBank EU639668-EU639685 。结论：前一种方法克隆扩增RGA 基因片段的效率很低，而后一种方法克隆扩增效果更
好，这为闽花6 号花生的遗传改良提供了理论基础。 |
| 英文摘要: |
| Objective: RGA fragments were cloned to investigate resistance and provide theoretical guidance for breeding in
peanut. Methods: In this study, degenerate PCR primers firstly designed to bind to DNA regions encoding conserved motifs within NBS
(nucleotide binding site) were used to amplify NBS-encoding regions from A. hypogaea L. Minhua Number 6. At the same time a pair of
degenerate primers was designed according to the result of multiple sequence alignment from known RGA sequences in peanut. RGA
fragments were amplified using homology cloning on the peanut variety Minhua number 6. Results: It was showed that for former there
was no sequence obtained in the study homologous to known sequences of RGA according to the sequencing results. While for later there
are 18 RGA sequences in 20 clones selected randomly manifested by BLASTN sequence analysis, GenBank numbers of which are
EU639668-EU639685. Conclusion: It shows that it is more efficiently to amplify corresponding RGA fragments by means of the RGA
sequence known to this species, which provides a shortcut for following RGA study and use. For that matter it provides theory basis for
genetic improvement in Minhua Number 6. |
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