|本期目录/Table of Contents|

中华鳖实时定量PCR内参基因的筛选与评价(PDF)

《内蒙古大学学报(自然科学版)》[ISSN:1000-9035/CN:22-1262/O4]

期数:
2017年05期
页码:
531-537
栏目:
研究论文
出版日期:
2017-09-15

文章信息/Info

Title:
Screening and Evaluation of qPCR Reference Genes in Pelodiscus sinensis
作者:
刘文婷1张贝贝1张耀红2管越强1
1河北大学生命科学学院,河北 保定 071002;
2.保定水产技术推广站,河北 保定 071051
Author(s):
LIU Wen-ting1ZHANG Bei-bei1ZHANG Yao-hong2GUAN Yue-qiang1
1.College of Life Sciences,Hebei University,Baoding 071002,China;
2.Baoding Fishery Technology Extension Station,Baoding 071051,China
关键词:
中华鳖实时定量PCR内参基因表达稳定性18S rRNA
Keywords:
Pelodiscus sinensisReal-time quantitative PCRreference genegene expression stability18S rRNA
分类号:
-
DOI:
-
文献标识码:
A
摘要:
选择合适的内参基因是采用q-PCR方法研究基因表达的前提.选取中华鳖GAPDH、EF1α、18S rRNA、Tubulin和β-actin等5个候选内参基因,通过q-PCR方法得到各基因在不同组织中的Ct值,利用4种内参筛选软件和综合评定法进行评价.geNorm和NormFinder软件分析均显示18S rRNA>EF1α>β-actin>GAPDH>Tubulin(稳定性由高到低).BestKeeper软件分析显示18S rRNA>GAPDH>EF1α>β-actin>Tubulin.RefFinder软件分析和综合评价法显示18S rRNA最佳.推荐优先使用18S rRNA或EF1α,不建议用β-actin和Tubulin.上述结果可为中华鳖和其它爬行动物的基因表达研究提供参考依据.
Abstract:
Selection of reference genes was important to the study of gene expression via qPCR method.GAPDH,EF1α,18S rRNA,Tubulin and β-actin of Chinese softshell turtle Pelodiscus sinensis were selected as the candidate reference genes.Cycle threshold value (Ct ) of different tissues were obtained through qPCR method,and the Ct value were further analyzed via 4 softwares and the comprehensive evaluation method on reference gene evaluation.The stability result of both geNorm and NormFinder was 18S rRNA> EF1α> β-actin> GAPDH> Tubulin (From high to low).The result of BestKeeper was 18S rRNA>GAPDH>EF1α>β-actin>Tubulin.The result of RefFinder and comprehensive evaluation method showed that 18S rRNA was the best reference gene.Therefore,18S rRNA or EF1α was recommend as the reference gene of P.sinensis,instead of β-actin and Tubulin.The study could provide a guidance and reference for the studying of gene expression of Chinese softshell turtle and other related reptiles.

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备注/Memo

备注/Memo:
收稿日期:2017-01-12;修回日期:2017-04-08
 基金项目:河北省自然科学基金项目(C2016201207);河北省现代农业产业技术体系淡水养殖创新团队项目
 作者简介:刘文婷(1990-),女,安徽肥西人,2015级硕士研究生.E-mail:280408479@qq.com
 通信作者:管越强(1973-),男,河北深泽人,教授,博士,主要研究方向:水生动物营养与免疫.E-mail:1426549783@qq.com
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