Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development
Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development
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DOI：
10.1046/j.1365-313x.1996.9050745.x
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发表时间：
1996-05-01
期刊：
影响因子：
7.2
通讯作者：
Visser, RGF
中科院分区：
文献类型：
--
作者：
Bachem, CWB;vanderHoeven, RS;Visser, RGF
利用高度同步的离体马铃薯马铃薯生根系统，结合扩增限制性片段多态性(AFLP匕首衍生的RNA指纹图谱技术（cDNA-AFLP双匕首），分析了马铃薯生根时间点及其周围的转录变化。对马铃薯主要贮藏蛋白patatin的特异转录本和淀粉生物合成途径关键基因adp -葡萄糖焦磷酸化酶的第二转录本进行了靶向表达分析。本文证实，cDNA-AFLP分析揭示的表达动力学与Northern分析的结果相当。此外，本文还报道了两个编码脂氧化酶的块茎特异性转录衍生片段（TDFs）的分离和分析，这两个片段在块茎形成的时间点周围被不同地诱导。对两个lox tdf的分析表明，有可能解剖单个转录本的表达方式，而Northern分析无法独立检测到。最后表明，利用dna - aflp可以实现快速、简单的条带身份验证。结果表明，dna - aflp是一种广泛适用的发育调控基因鉴定技术。
Using a highly synchronous in vitro tuberization system, in combination with an amplified restriction fragment polymorphism (AFLP dagger-derived technique for RNA fingerprinting (cDNA-AFLP double dagger), transcriptional changes at and around the time point of potato tuberization have been analyzed. The targeted expression analysis of a specific transcript coding for the major potato storage protein, patatin and a second transcript, coding for ADP-glucose pyrophosphorylase, a key gene in the starch biosynthetic pathway is described. This paper confirms that kinetics of expression revealed by cDNA-AFLP analysis are comparable to those found in Northern analysis. Furthermore, this paper reports the isolation and analysis of two tuber-specific transcript-derived fragments TDFs) coding for the lipoxygenase enzyme, which are differentially induced around the time point of tuber formation. Analysis of the two lox TDFs demonstrates that it is possible to dissect the expression modalities of individual transcripts, not independently detectable by Northern analysis. Finally, it is shown that using cDNA-AFLP, rapid and simple verification of band identity may be achieved. The results indicate that cDNA-AFLP is a broadly applicable technology for identifying developmentally regulated genes.