干旱胁迫下木薯AP2转录因子的荧光定量PCR分析

为研究木薯的两个A P2基因在干旱胁迫下的表达模式,对木薯品种华南5号进行干旱胁迫处理,提取各个时间点离区部位的RNA,以其反转录的cDNA为模板,应用SYBR Green Ⅰ荧光定量PCR检测AP2转录因子基因的差异性表达.结果表明,目的基因的熔解曲线都只出现一个峰,说明引物特异性强,且各基因的扩增效率与参照基因A ctin接近,试验结果可用2^-△△CT计算法进行分析.经分析,两个AP2家族成员在干旱胁迫下均诱导表达,其表达模式不完全相同且存在一定的相关性,这与之前做过的基因芯片杂交结果相符....

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Bibliographic Details
Published in广东农业科学 Vol. 41; no. 10; pp. 126 - 131
Main Author 李雅韵 廖文彬 王斌 杨义伶 郭鑫 杨子 彭明
Format Journal Article
LanguageChinese
Published 海南大学农学院,海南海口570228 2014
中国热带农业科学院热带生物技术研究所,海南海口571101%中国热带农业科学院热带生物技术研究所,海南海口,571101
Subjects
AP2转录因子
基因表达
干旱胁迫
荧光定量PCR
AP2转录因子
基因表达
干旱胁迫
荧光定量PCR
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Summary:为研究木薯的两个A P2基因在干旱胁迫下的表达模式,对木薯品种华南5号进行干旱胁迫处理,提取各个时间点离区部位的RNA,以其反转录的cDNA为模板,应用SYBR Green Ⅰ荧光定量PCR检测AP2转录因子基因的差异性表达.结果表明,目的基因的熔解曲线都只出现一个峰,说明引物特异性强,且各基因的扩增效率与参照基因A ctin接近,试验结果可用2^-△△CT计算法进行分析.经分析,两个AP2家族成员在干旱胁迫下均诱导表达,其表达模式不完全相同且存在一定的相关性,这与之前做过的基因芯片杂交结果相符.
Bibliography:44-1267/S
In order to study the expression patterns of two AP2 genes in cassava under drought stress, the cassava variety SC5 was treated with drought stress and RNA was extracted from the leaf abscission zone of different time points. We detected the different expression of AP2 transcription factors by SYBR Green I real-time fluorescent quantitative PCR with the cDNA obtained from reverse transcription as templates. The results showed that melting curves of both target genes had only one peak, indicating that the primers were specific and the amplification efficiency of target genes were close to reference gene A ctin. The experimental results could be analyzed by 2^-△△CT calculation method. After analysis, the result showed that the two AP2 family members were induced under drought stress, the expression patterns were not identical and there was certain correlation. These results were consistent with microarray hybridization results.
drought stress; AP2 transcription factor; real-time fluorescent quantitative
ISSN:1004-874X