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Reference gene selection for microRNA expression analysis in the Pacific oyster
Huang, Wen1,2,3,4; Xu, Fei1,3,4; Wang, Wei1,3,4; Li, Li1,2,4; Que, Huayong1,2,3,4; Zhang, Guofan1,2,3,4
2018-09-01
发表期刊ROMANIAN BIOTECHNOLOGICAL LETTERS
ISSN1224-5984
卷号23期号:5页码:14043-14050
摘要Real-time quantitative polymerase chain reaction (qRT-PCR) is a widely used technique for microRNA (miRNA) expression analysis. The accuracy and reproducibility of this technique depends on the availability of stably expressed reference genes, which is absent in molluscs. Here, based on the microRNA transcriptome data of the Pacific oyster, Crassostrea gigas, candidate reference genes were selected from the stably expressed microRNAs and the commonly used housekeeping genes (U1, U6, and 5.8S). The stability of expression was evaluated and the comprehensive ranking of the reference genes was obtained using two statistical algorithms, geNorm and NormFinder. Specific combinations of reference genes were recommended for the developmental larval samples (DLS) and adult organ samples (AOS), respectively; a combination of cgi-miR-71 and Ul was recommended for DLS and a combination of cgi-let-7, cgi-miR-87, and Ul for AOS, under normal physiological conditions. The effectiveness of the reference gene combinations was further validated by quantitative analysis of two selected miRNAs in the DLS and AOS, respectively. This work was the first systematic analysis of miRNA reference gene selection in molluscs. The reference genes identified in this study will be useful for the functional analysis of miRNA in Crassostrea gigas, and also for the miRNA reference gene selection in other molluscan species.
关键词Crassostrea gigas MicroRNA Real-time quantitative PCR Reference gene
DOI10.26327/RBL2018.224
收录类别SCI
语种英语
资助项目Taishan Scholars Climbing Program of Shandong ; China Agriculture Research System[CARS-49] ; Guangxi Natural Science Foundation[2016GXNSFBA380028] ; National Natural Science Foundation of China[31372515] ; National Natural Science Foundation of China[31372515] ; Guangxi Natural Science Foundation[2016GXNSFBA380028] ; China Agriculture Research System[CARS-49] ; Taishan Scholars Climbing Program of Shandong
WOS研究方向Biotechnology & Applied Microbiology
WOS类目Biotechnology & Applied Microbiology
WOS记录号WOS:000454870500017
出版者ARS DOCENDI
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文献类型期刊论文
版本出版稿
条目标识符http://ir.qdio.ac.cn/handle/337002/155956
专题实验海洋生物学重点实验室
通讯作者Que, Huayong; Zhang, Guofan
作者单位1.Chinese Acad Sci, Inst Oceanol, Key Lab Expt Marine Biol, Qingdao, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China
3.Qingdao Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao, Peoples R China
4.Chinese Acad Sci, Ctr Ocean Mega Sci, Beijing, Peoples R China
第一作者单位中国科学院海洋研究所
通讯作者单位中国科学院海洋研究所
推荐引用方式
GB/T 7714
Huang, Wen,Xu, Fei,Wang, Wei,et al. Reference gene selection for microRNA expression analysis in the Pacific oyster[J]. ROMANIAN BIOTECHNOLOGICAL LETTERS,2018,23(5):14043-14050.
APA Huang, Wen,Xu, Fei,Wang, Wei,Li, Li,Que, Huayong,&Zhang, Guofan.(2018).Reference gene selection for microRNA expression analysis in the Pacific oyster.ROMANIAN BIOTECHNOLOGICAL LETTERS,23(5),14043-14050.
MLA Huang, Wen,et al."Reference gene selection for microRNA expression analysis in the Pacific oyster".ROMANIAN BIOTECHNOLOGICAL LETTERS 23.5(2018):14043-14050.
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