摘要
利用基因克隆和测序,从‘妃子笑’荔枝果皮中获得了β-Actin、GAPDH和18S rRNA 3个qPCR分析常用的内参基因全长序列,其长度分别为1 273、1 368和1 712 bp;3个基因与其他物种高度同源,氨基酸序列相似性均超过了98%.在此基础上,结合已报道的UBQ、eEF和25S rRNA 3个常用的内参基因,对β-Actin、GAPDH、18S rRNA,UBQ、eEF和25S rRNA 6个常用内参基因在荔枝果实发育不同阶段和外源生长调节剂处理后表达稳定性进行了分析,同时比较了geNorm、Norm Finder和BestKeeper 3种不同算法的差异.结果表明:以上6个基因中,β-Actin基因在3种不同算法下均保持了较好的表达稳定性.
The full-length sequences of β-Actin, GAPDH and 18S rRNA which are frequently used as ref- erence genes in qPCR analysis were obtained from the pericarp of litchi （Litchi chinensis Sonn. cv. Feizixiao） by gene cloning and sequencing. Their sequence lengths were 1 273,1 368 and 1 712 bp, respectively. Sequence analysis showed that the cloned sequences had very high similarity with other species. Furthermore, the stability of six genes, including β-Actin, glyceraldehyde-3-phosphate dehydrogenase （ GAPDH）, 18S rRNA, ubiquitin （ UBQ）, elongation factor （eEF） and 25S rRNA, was determined in litchi pericarp under different developmental stages and regulator treatments in this study. Three different analytic calculation methods including geNorm, NormFinder and BestKeeper were compared. Among the six genes tested,β-Actin appeared to be the most stable single gene under three different calculation methods.
出处
《华南农业大学学报》
CAS
CSCD
北大核心
2012年第3期301-306,共6页
Journal of South China Agricultural University
基金
国家自然科学基金(30971985)
国家科技支撑计划项目(2006BAD 0lAl705)
现代农业产业技术体系建设专项资金(CARS-33)
关键词
荔枝
qPCR
内参基因
克隆
稳定性分析
Litchi chinensis
qPCR
reference genes
cloning
stability analysis
