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    魏永赞, 赖彪, 胡福初, 李晓静, 胡桂兵, 王惠聪. 用于荔枝qPCR分析的内参基因克隆及稳定性分析[J]. 华南农业大学学报, 2012, 33(3): 301-306. DOI: 10.7671/j.issn.1001-411X.2012.03.006
    引用本文: 魏永赞, 赖彪, 胡福初, 李晓静, 胡桂兵, 王惠聪. 用于荔枝qPCR分析的内参基因克隆及稳定性分析[J]. 华南农业大学学报, 2012, 33(3): 301-306. DOI: 10.7671/j.issn.1001-411X.2012.03.006
    shu
    WEI Yong-zan, LAI Biao, HU Fu-chu, LI Xiao-jing, HU Gui-bing, WANG Hui-cong. Cloning and Stability Analysis of Reference Genes for Expression Studies by Quantitative Real-Time PCR in Litchi[J]. Journal of South China Agricultural University, 2012, 33(3): 301-306. DOI: 10.7671/j.issn.1001-411X.2012.03.006
    Citation: WEI Yong-zan, LAI Biao, HU Fu-chu, LI Xiao-jing, HU Gui-bing, WANG Hui-cong. Cloning and Stability Analysis of Reference Genes for Expression Studies by Quantitative Real-Time PCR in Litchi[J]. Journal of South China Agricultural University, 2012, 33(3): 301-306. DOI: 10.7671/j.issn.1001-411X.2012.03.006
    shu

    用于荔枝qPCR分析的内参基因克隆及稳定性分析

    Cloning and Stability Analysis of Reference Genes for Expression Studies by Quantitative Real-Time PCR in Litchi

      摘要
    • 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、NormFinder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.

       

      Abstract: The full-length sequences of β-Actin,GAPDH and 18S rRNA which are frequently used as reference 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.

       

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