生物学杂志

生物学杂志

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蓝点马鲛鱼内参基因的克隆及表达稳定性评价

  

  1. 宁波大学 海洋学院， 宁波 315211
  • 出版日期:2016-06-18 发布日期:2016-06-18
  • 通讯作者: 薛良义，教授，博士生导师，主要从事鱼类分子生物学研究，E-mail：xueliangyi@nbu.edu.cn
  • 作者简介:张家玲，硕士研究生，主要从事海洋生物基因资源，E-mail：xin.jialing@163.com
  • 基金资助:
    宁波市科技局项目（编号：2012C10035）

Cloning and expression stability evaluation of reference genes in Spanish Mackerel (Scomberomorus niphonius)

  1. School of Marine Science, Ningbo University, Ningbo 315211, China
  • Online:2016-06-18 Published:2016-06-18

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摘要： 合适内参基因的选择是实时荧光定量PCR技术准确测定目的基因表达量的前提。首先克隆了蓝点马鲛鱼3个内参基因β-actin、GAPDH和EF1-α的部分序列，并通过实时荧光定量PCR分析β-actin、GAPDH、EF1-α和18S rRNA 4个内参基因在蓝点马鲛鱼各组织中的Ct值，应用3种内参基因筛选软件（geNorm，NormFinder和Bestkeeper）综合分析这4个基因的表达稳定性。结果表明最稳定的内参基因是EF1-α，其次是18S rRNA。研究结果为今后蓝点马鲛鱼合适内参基因的选择提供了依据。

关键词: 蓝点马鲛鱼, 实时荧光定量PCR, 内参基因, geNorm软件, 表达稳定性

Abstract: Selection of a suitable reference gene is an important prerequisite for precise analysis of target gene expression by Real-time quantitative PCR(qPCR). In this paper, the partial sequence of three reference genes, β-actin and GAPDH, EF1-α, from spanish mackerel Scomberomorus niphonius were firstly cloned, and then the Ct values of the four reference genes in various tissues were measured by qPCR. Three different statistical algorithms including geNorm, NormFinder and BestKeeper were used to analyze the expression stability of the four reference genes in different tissues. The result showed that the most stable reference gene was EF1-a, followed by 18S ribosomal RNA. It provides a useful basis for selecting of the appropriate reference gene in Spanish Mackerel.

Key words: Scomberomorus niphonius, qPCR, reference gene, geNorm, expression stability