印度谷螟实时荧光定量PCR内参基因的选择

发布时间：2018-06-07 06:22

本文选题：印度谷螟 + 仓储害虫　；参考：《中国科学:生命科学》2016年10期

【摘要】：实时荧光定量PCR(qRT-PCR)是研究基因表达的重要手段之一,而选择合适的内参基因是获得可靠研究结果的基础.本研究以世界性分布的重要仓储害虫印度谷螟(Plodia interpunctella)为研究对象,旨在筛选出该虫不同发育阶段及不同品系qRT-PCR的最适内参基因,并利用两个独立软件(ge Norm和Norm Finder)对8个备选内参基因的表达稳定性进行了评价.结果表明,不同发育阶段最稳定表达的内参基因为Ef1a,而不同品系最稳定的是b-Actin和Ef1a.利用筛选到的Ef1a,b-Actin及两者组合对目标基因(过氧化物酶POD)进行标准化,印度谷螟不同品系间的POD基因表达量存在显著差异,而以稳定性最差的SD作为内参时,不同品系间POD的相对表达量并无显著差异.这表明,在开展qRT-PCR研究时评价内参基因的稳定性非常必要,且基于筛选到的最佳基因作为内参时,才能准确获得目标基因的相对表达水平.这些结果将有助于更好地研究印度谷螟发育及品系特异相关基因的功能,并为筛选印度谷螟在其他条件下的最适内参基因提供参考.
[Abstract]:Real-time quantitative PCR qRT-PCR is one of the important methods to study gene expression, and the selection of suitable internal reference gene is the basis for obtaining reliable results. In this study, Plodia interpunctura, a worldwide important storage pest, was used to screen the optimal internal reference genes for different stages of development and different strains of qRT-PCR. The stability of the expression of 8 candidate reference genes was evaluated by using two independent softwares named ge Norm and Norm Finder. The results showed that the most stable endoparameters expressed in different developmental stages were Ef1a, while the most stable ones in different strains were b-Actin and Ef1a. The target gene (POD) was standardized by using the selected Ef1aPb-Actin and their combinations. There were significant differences in the expression of POD gene among different strains of Indian grain borer, but when SD with the lowest stability was used as the internal control. There was no significant difference in the relative expression of POD among different strains. This indicates that it is necessary to evaluate the stability of the internal reference gene in the qRT-PCR study, and the relative expression level of the target gene can be accurately obtained only when the best selected gene is used as the internal reference. These results will be helpful to study the development of Indian grain borer and the function of strain specific genes, and provide a reference for screening the most suitable internal reference genes of Indian grain borer under other conditions.
【作者单位】：南京财经大学食品科学与工程学院江苏省现代粮食流通与安全协同创新中心;南京大学生命科学学院演化发育生物学研究所;兰州大学草地农业科技学院草地农业生态系统国家重点实验室;
【基金】：粮食公益性行业科研专项(批准号:201413007-2) 国家科技支撑计划(批准号:2013BAD17B01) 国家自然科学基金(批准号:31000828) 江苏省高校优势学科建设工程资助
【分类号】：S379.5

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