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核农学报  2011 

文心兰cDNA-AFLP体系的建立及TDFs分析

, PP. 1142-1147

Keywords: 文心兰,cDNA-AFLP,引物筛选,TDFs

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Abstract:

通过总RNA提取、cDNA合成等试验,建立cDNA-AFLP分析体系。结果表明,采用MseⅠ和EcoRⅠ酶切3h,预扩增产物稀释15倍作为选择性扩增模板,利用筛选的引物进行扩增,可获得丰度和多态性较高的指纹图谱。利用59对引物获得约1160条TDFs(Transcription-derivedfragments)。回收、克隆其中128条进行测序,获得102条可读序列,GenBank登陆号为JG014357~JG014458。BLASTN/X同源比对发现,87%的条带在NCBI数据库中能找到功能已知的同源序列;其中2条TDFs分别可能与文心兰子房发育(TDF0073a)和花器官挥发性香味物质合成(TDF0139a)相关。序列分析表明TDFs对应基因的功能涉及植物生命活动的诸多领域。cDNA-AFLP分析体系的建立及TDFs序列分析为进一步克隆花发育相关基因,研究文心兰花发育相关分子调控机制奠定了基础。

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