家兔不同发育阶段和组织中内参基因的稳定性分析

doi:10.11843/j.issn.0366-6964.2016.03.008

摘要/Abstract

摘要：

本研究旨在筛选出新西兰白兔不同时期不同器官中表达最稳定的内参基因。以胚胎期(E28.5)、幼龄期(10 d)和成年期(A)3个阶段的新西兰白兔的肾、心和肝3种组织作为研究对象，按照RT-qPCR试验最低要求试验信息量(MIQE)的指导，运用RT-qPCR和在线内参基因稳定性评价工具RefFinder，比较6个常用的内参基因（GAPDH、HPRT1、18S rRNA、B2M、Sdha、β-actin）mRNA的表达稳定性。结果表明，新西兰白兔的肝和肾组织中的6个内参基因在3个发育阶段最稳定的3个内参基因依次是β-actin、HPRT1、GAPDH，而在心组织中最稳定的3个内参基因依次是β-actin、HPRT1、Sdha。在新西兰白兔胚胎期的3种组织中，最稳定的3个内参基因依次是GAPDH、β-actin、HPRT1，而在幼龄期和成年期最稳定的3个内参基因依次是HPRT1、GAPDH、β-actin。对新西兰白兔的3个发育阶段和3种组织的内参基因稳定性结果分析表明，最稳定的3个内参基因依次是β-actin、HPRT1、GAPDH。本研究证实，新西兰白兔不同的发育时期和组织中，最稳定的内参基因不同。推荐在进行新西兰白兔的相关试验时，可以选择上述表达最稳定的至少3个内参基因的几何平均数作为标准化相关RT-qPCR数据的指标，以确保试验组个体间的差异最小，和小变量的准确数据分析。

关键词: 内参基因, 表达稳定性, MIQE, RefFinder

Abstract:

The purpose of this study was to screen the most stable reference genes in different development periods and tissues in New Zealand White rabbit.Quantitative real-time Polymerase Chain Reaction(RT-qPCR)，a latest online reference genes stability assessment tool RefFinder and Minimum Information for Publication of Quantitative Real-time PCR Experiments(MIQE) guideline were used to compare the expression levels of 6 commonly used reference genes(GAPDH，HPRT1，18S rRNA，B2M，Sdha，β-actin) mRNA in 3 tissues(heart，liver，kidney) of rabbit at embryo(E28.5)，juvenile(10 d) and adult stages(A).The result showed that the most stable reference genes in liver and kidney of New Zealand White rabbit in 3 development periods were β-actin， HPRT1 and GAPDH，successively.But for heart，they were β-actin，HPRT1， Sdha.The most stable reference genes in embryo period of New Zealand White rabbit in 3 tissues were GAPDH，β-actin and HPRT1，successively.For juvenile and adult stages，they were HPRT1，GAPDH and β-actin. To all 3 periods and all 3 tissues of New Zealand White rabbit，the most stable reference genes were successively β-actin，HPRT1 and GAPDH.This study confirmed that the most stable reference genes were different in different periods and tissues of New Zealand White rabbit.To ensure minimal variation between individuals in an experimental group and the accurate statistical analysis of small fold changes，we recommend normalizing RT-qPCR data to the geometric mean of at least 3 validated reference genes as above when related New Zealand White rabbit trials were performed.

Key words: reference genes, expression stability, MIQE, RefFinder

中图分类号:

陈瑞，杨晓农，曾婉秋，文娟. 家兔不同发育阶段和组织中内参基因的稳定性分析[J]. 畜牧兽医学报, 2016, 47(3): 477-483.

CHEN Rui，YANG Xiao-nong，ZENG Wan-qiu，WEN Juan. Expression Stability Analysis of Reference Genes in Different Development Periods and Tissues in Oryctolagus Cuniculus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(3): 477-483.

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