苹果实时荧光定量PCR分析中内参基因的筛选Screening of reference genes for Real-time Fluorescence Quantitative PCR in apple(Malus×domestica)
周兰,张利义,张彩霞,康国栋,田义,丛佩华
摘要(Abstract)：
【目的】为筛选苹果实时荧光定量PCR实验中最适内参基因,【方法】应用实时荧光定量PCR技术,分析5个传统内参基因18SrRNA、ACTB、GAPDH、UBQ、TUB在苹果不同基因型、不同组织、果实不同发育时期的mRNA表达差异情况。供试的6个不同基因型苹果分别为:新疆野生苹果、八棱海棠、丽江山荆子、‘津轻’、‘国庆’、‘金冠’;6种不同组织为果皮、果肉、叶片、愈伤组织、花瓣、种子;5个果实发育不同时期为花后28、50、74、95、115 d。【结果】经geNorm程序分析发现5种内参基因的表达稳定性各异,UBQ在果实不同基因型和不同发育时期的基因表达分析中最稳定;ACTB和UBQ在6种不同组织中表达均稳定。【结论】UBQ在参试样品中表达均比较稳定,是研究苹果基因表达分析中理想的内参基因。
关键词(KeyWords)： 苹果;实时荧光定量PCR;内参基因;geNorm程序
基金项目(Foundation): 现代苹果产业技术体系(nycytx-08-01);; 国家863项目(2011AA10020404)
作者(Author): 周兰,张利义,张彩霞,康国栋,田义,丛佩华
DOI: 10.13925/j.cnki.gsxb.2012.06.008
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