• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>BMC Molecular Biology >Evaluation of endogenous references for gene expression profiling in different tissues of the oriental fruit fly Bactrocera dorsalis (Diptera: Tephritidae)
【24h】

Evaluation of endogenous references for gene expression profiling in different tissues of the oriental fruit fly Bactrocera dorsalis (Diptera: Tephritidae)

机译：评价东方果蝇小实蝇（Bactrocera dorsalis）（双翅目：蝇科）不同组织中基因表达谱的内源性参考

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Background Quantitative real-time reverse transcriptase PCR (RT-qPCR) has been widely used for quantification of mRNA as a way to determine key genes involved in different biological processes. For accurate gene quantification analysis, normalization of RT-qPCR data is absolutely essential. To date, normalization is most frequently achieved by the use of internal controls, often referred to as reference genes. However, several studies have shown that the reference genes used for the quantification of mRNA expression can be affected by the experimental set-up or cell type resulting in variation of the expression level of these key genes. Therefore, the evaluation of reference genes is critical for gene expression profiling, which is often neglected in gene expression studies of insects. For this purpose, ten candidate reference genes were investigated in three different tissues (midgut, Malpighian tubules, and fat body) of the oriental fruit fly, Bactrocera dorsalis (Hendel). Results Two different programs, geNorm and Normfinder, were used to analyze the data. According to geNorm, α-TUB + ACT5 are the most appropriate reference genes for gene expression profiling across the three different tissues in the female flies, while ACT3 + α-TUB are considered as the best for males. Furthermore, we evaluated the stability of the candidate reference genes to determine the sexual differences in the same tissue. In the midgut and Malpighian tubules, ACT2 + α-TUB are the best choice for both males and females. However, α-TUB + ACT1 are the best pair for fat body. Meanwhile, the results calculated by Normfinder are quite the same as the results with geNorm; α-TUB is always one of the most stable genes in each sample validated by the two programs. Conclusions In this study, we validated the suitable reference genes for gene expression profiling in different tissues of B. dorsalis. Moreover, appropriate reference genes were selected out for gene expression profiling of the same tissues taking the sexual differences into consideration. This work not only formed a solid basis for future gene expression study in B. dorsalis, but also will serve as a resource to screen reference genes for gene expression studies in any other insects.
机译：背景技术实时定量逆转录酶PCR（RT-qPCR）已被广泛用于mRNA定量，作为一种确定涉及不同生物学过程的关键基因的方法。对于精确的基因定量分析，RT-qPCR数据的标准化绝对必要。迄今为止，最常通过使用内部对照（通常称为参考基因）来实现标准化。但是，一些研究表明，用于定量mRNA表达的参考基因可能会受到实验设置或细胞类型的影响，从而导致这些关键基因的表达水平发生变化。因此，参考基因的评估对于基因表达谱分析至关重要，而在昆虫的基因表达研究中常常忽略了这一点。为此，在东方实蝇Bactrocera dorsalis（Hendel）的三个不同组织（中肠，马尔皮基小管和脂肪体）中研究了十个候选参考基因。结果使用两个不同的程序geNorm和Normfinder来分析数据。根据geNorm的研究，α-TUB+ ACT5是在雌蝇的三个不同组织中进行基因表达谱分析的最合适参考基因，而ACT3 +α-TUB被认为是最适合雄性的基因。此外，我们评估了候选参考基因的稳定性，以确定同一组织中的性别差异。在中肠和Malpighian小管中，ACT2 +α-TUB是男性和女性的最佳选择。但是，α-TUB+ ACT1是最适合肥胖的身体。同时，Normfinder计算的结果与geNorm的结果完全相同。 α-TUB始终是通过两个程序验证的每个样品中最稳定的基因之一。结论在这项研究中，我们验证了适合在背侧双歧杆菌不同组织中进行基因表达谱分析的参考基因。此外，考虑到性别差异，选择适当的参考基因用于相同组织的基因表达谱分析。这项工作不仅为今后在桔小实蝇中进行基因表达研究奠定了坚实的基础，而且还将成为筛选参考基因以在其他任何昆虫中进行基因表达研究的资源。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号