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Stably Expressed Housekeeping Genes across Developmental Stages in the Two-Spotted Spider Mite, Tetranychus urticae

文献类型： 外文期刊

作者： Yang, Chunxiao 2 ; Pan, Huipeng 2 ; Liu, Yong 1 ; Zhou, Xuguo 2 ;

作者机构： 1.Hunan Acad Agr Sci, Inst Plant Protect, Changsha, Hunan, Peoples R China

2.Univ Kentucky, Dept Entomol, Lexington, KY 40546 USA

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2015 年 10 卷 3 期

页码：

收录情况： SCI

摘要： Quantitative real-time PCR (qRT-PCR) is a reliable and reproducible technique for measuring mRNA expression. To facilitate gene expression studies and obtain more accurate qRT-PCR analysis, normalization relative to stable housekeeping genes is mandatory. In this study, ten housekeeping genes, including beta-actin (Actin), elongation factor 1 alpha (EF1A), glyceralde hyde-3-phosphate dehydrogenase (GAPDH), ribosomal protein L13 (RPL13), ribosomal protein 49 (RP49), alpha-tubulin (Tubulin), vacuolar-type H+-ATPase (v-ATPase), succinate dehydrogenase subunit A (SDHA), 28S ribosomal RNA (28S), and 18S ribosomal RNA (18S) from the two-spotted spider mite, Tetranychus urticae, were selected as the candidate reference genes. Four algorithms, geNorm, Normfinder, BestKeeper, and the Delta C-t method, were used to evaluate the performance of these candidates as endogenous controls across different developmental stages. In addition, RefFinder, which integrates the above-mentioned software tools, provided the overall ranking of the stability/suitability of these candidate reference genes. Among them, PRL13 and v-ATPase were the two most stable housekeeping genes across different developmental stages. This work is the first step toward establishing a standardized qRT-PCR analysis in T. urticae following the MIQE guideline. With the recent release of the T. urticae genome, results from this study provide a critical piece for the subsequent genomics and functional genomics research in this emerging model system.

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