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Characterizing the expression of translation elongation factor gene EF1 alpha in pear (Pyrus) fruit: evaluation of EF1 alpha as a housekeeping gene

文献类型： 外文期刊

作者： Wang, Yue-zhi 1 ; Dai, Mei-song 1 ; Cai, Dan-ying 1 ; Miao, Lixiang 1 ; Wei, Lingzhu 1 ; Shi, Ze-bin 1 ;

作者机构： 1.Zhejiang Acad Agr Sci, Inst Hort, Hangzhou 310021, Zhejiang, Peoples R China

关键词： Gene expression; mRNA sequencing; qRT-PCR; Reference gene

期刊名称：TREE GENETICS & GENOMES （ 影响因子：2.297； 五年影响因子：2.547 ）

ISSN： 1614-2942

年卷期： 2018 年 14 卷 4 期

页码：

收录情况： SCI

摘要： Reference genes are a key factor for the sensitivity and reliability of quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. The elongation factor gene EF1 alpha encodes a highly conserved ubiquitous protein that functions in the binding of aminoacyl-tRNA to the ribosome during peptide synthesis in eukaryotes, which is proposed for qRT-PCR as an internal reference for its putative housekeeping gene character. However, information about the paralogous copies and expression diversification of EF1 alpha has been neglected and ill defined. In this study, the members of pear (Pyrus) EF1 alpha were explored by mining public data. The family- and/or species-specific members of EF1 alpha in Rosaceae plants or other species revealed by phylogenetic analysis suggested that the specific gene expansion occurred after the family or species diversification. The expression analysis based on the high-throughput sequencing and sequence differences typing provided a high resolution to distinguish the expression among the members of pear EF1 alpha. The EF1 alpha members showed an obviously unstable expression in both pear and apple (Malus x domestica) fruits at different developmental stages, clustered by two conserved expression patterns in pear fruit. The complementary expression among certain pear EF1 alpha members leads to the sum expression of these members having a higher level of expression stability among the fruit at different developmental stages, which can be used as an approach to optimize EF1 alpha as the internal reference. The result can also provide insight into the expression characteristics of pear EF1 alpha in other tissues of pear. In addition, it indicates that any two genes having complementary expression patterns can be used as a reference for qPCR analysis with the high stability of their mean expression value.

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