摘要
实时荧光定量PCR 是由美国Applied Biosystems 公司于1996年在传统的PCR 技术基础上发展起来的一种新的核酸定量技术（ Mackay，2004），具有定量准确、灵敏度高、特异性强、重复性好及高通量等优点（Huggett et al．，2005），已成为医学、农学、林学、微生物学等众多领域中进行基因表达和转录组分析的常用技术之一（ Postollec et al．，2011；袁亚男等，2008）。
The real-time PCR is an important method for analyzing gene expression levels. Selection of reference genes suitable for calibration of the expression of objective gene according to specific experimental materials and conditions is a prerequisite for obtaining reliable RT-PCR results. In this study,we used different individuals and different organs of black locust（Robinia pseudoacacia）to test the stability of six internal genes such as Actin gene （ACT）,18S ribosomal RNA （18S rRNA）,adenosylmethionine decarboxylase gene （SAMDC）,helicase gene （Helicase）,elongation factor gene （EF1-α）,glyceraldehyde-3-phosphate-dehydrogenase gene （GAPDH） in the real-time PCR. After being analyzed with Genorm and NormFinder,the results showed that the ACT and 18S rRNA gene were the most stable genes by using different plant individuals as experimental material,while the ACT and GAPDH were the most stable genes with using different tissues and organs. Since some controversies existed with the 18S rRNA as reference gene,the both ACT and GAPDH used for reference gene in various test conditions obtained more accurate results. This study has important implications in obtaining a more accurate result of quantitative expression of black locust.
出处
《林业科学》
EI
CAS
CSCD
北大核心
2014年第9期167-172,共6页
Scientia Silvae Sinicae
基金
国家自然科学基金项目“刺槐杂交结实率低的机理研究”(31170629)
国家科技支撑项目“抗逆生态树种刺槐新品种选育技术研究”(2012BAD01B0601)
北京市科委项目“首都平原百万亩造林科技支撑工程”(Z121100008512002)
