Reference gene selection for real-time RT-PCR normalization in rice field eel (Monopterus albus) during gonad development

被引:52
作者
Hu, Qing [1 ,2 ,3 ]
Guo, Wei [1 ,2 ,3 ]
Gao, Yu [1 ,2 ,3 ]
Tang, Rong [1 ,2 ,3 ]
Li, Dapeng [1 ,2 ,3 ,4 ]
机构
[1] Huazhong Agr Univ, Coll Fisheries, Wuhan 430070, Peoples R China
[2] Freshwater Aquaculture Collaborat Innovat Ctr Hub, Wuhan 430070, Peoples R China
[3] Minist Agr, Key Lab Freshwater Anim Breeding, Wuhan 430070, Peoples R China
[4] Hunan Univ Arts & Sci, Life Sci Coll, Changde 415000, Peoples R China
关键词
Reference gene; Real-time RT-PCR; Stability; Monopterus albus; POTENTIAL REFERENCE GENES; POLYMERASE-CHAIN-REACTION; HOUSEKEEPING GENES; ATLANTIC SALMON; INTERNAL CONTROL; EXPRESSION; VALIDATION; TISSUES; ZEBRAFISH; ASSAYS;
D O I
10.1007/s10695-014-9962-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Real-time reverse transcriptase (RT) polymerase chain reaction (PCR) requires data normalization using an appropriate reference gene in order to obtain more reliable results with biological significance. We cloned a partial sequence of elongation factor-1-alpha (EF1 alpha) and ribosomal protein L17 (RPL17) from Monopterus albus. We investigated the suitability of five commonly used reference genes [18S ribosomal RNA (18S), cytoskeletal protein (beta-actin), glyceraldehyde phosphate dehydrogenase (GAPDH), EF1 alpha and RPL17] as potential quantitative reference genes for normalizing real-time RT-PCR data generated in gonads of different developmental stages and in other tissues of M. albus. Analysis of the data indicated that 18S, beta-actin and GAPDH are not suitable as reference genes because of their levels of variations of expression. EF1 alpha and RPL17 might be suitable as reference genes in the gonads of different developmental stages as well as in other tissues of M. albus.
引用
收藏
页码:1721 / 1730
页数:10
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