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Fine Mapping of AST Gene in Arabidopsis 被引量:1
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作者 毛爱军 王台 宋艳茹 《Acta Botanica Sinica》 CSCD 2003年第1期88-92,共5页
The ast ( anthocyanin spotted testa) mutant, which was induced by carbon ion radiation, was a single recessive gene mutant of Arabidopsis thaliana (L.) Heynh. with spotted pigment in seed coats, and involved in the an... The ast ( anthocyanin spotted testa) mutant, which was induced by carbon ion radiation, was a single recessive gene mutant of Arabidopsis thaliana (L.) Heynh. with spotted pigment in seed coats, and involved in the anthocyanin biosynthesis. To clone the AST gene by map-based cloning strategy, a series of molecular markers were designed according to the SNPs (single nucleotide polymophisms) and insertion/deletion polymophisms in the Arabidopsis database. With these molecular markers, the fine-structure mapping of the AST gene was finished, the AST locus was located in BAC clone T13M11. It was suggested that the AST candidate gene was T13M11. 8 in the T13M11 This gene was 1432 bp long with 6 exons and 5 introns. The putative protein of T13M11. 8 gene was similar to dihydroflavonol 4-reductase (DFR), which was an important enzyme in the anthocyanin biosynthesis pathway. 展开更多
关键词 ast mutant ast gene molecular marker fine mapping map-based cloning ARABIDOPSIS
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用差异显示反转录PCR银染技术研究植物基因表达的差异 被引量:7
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作者 毛爱军 王台 宋艳茹 《生物化学与生物物理进展》 SCIE CAS CSCD 北大核心 2001年第5期736-739,共4页
通过调整差异显示反转录PCR (DDRT PCR)中总RNA、锚定引物、随机引物、cDNA和dNTP等关键试剂的用量 ,优化了适用于银染检测的DDRT PCR方法 .PCR扩增产物经 6 %变性聚丙烯酰胺凝胶垂直电泳分离后 ,银染能检测到多而清晰的条带 .泳道中的... 通过调整差异显示反转录PCR (DDRT PCR)中总RNA、锚定引物、随机引物、cDNA和dNTP等关键试剂的用量 ,优化了适用于银染检测的DDRT PCR方法 .PCR扩增产物经 6 %变性聚丙烯酰胺凝胶垂直电泳分离后 ,银染能检测到多而清晰的条带 .泳道中的条带数最少为 40个 ,最多达 80个 ,平均为 6 0个 ,条带大小分布在 10 0~ 90 0bp范围 ,灵敏度为 5pg/mm2 .此方法操作简便快速 ,灵敏度高 ,重复性好 .采用这个改良的方法 ,分析了拟南芥野生型和ast突变型基因表达的差异 .从 16 0 0 0个cDNA扩增产物条带中筛选出 2 8个差异条带 .二次PCR扩增后 ,进一步筛选出 13个差异条带 ,其中 7个是野生型特异表达的 ,6个是突变型特异表达的 。 展开更多
关键词 差异显示反转录PCR 银染 拟南芥 ast突变 植物 基因表达
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