Identification and evaluation of reference genes for expression studies by RT-qPCR during embryonic development of the emerging model organism, Macrobrachium olfersii

被引：29

作者：

Jaramillo, Michael L. ^{[1
]}

Ammar, Dib ^{[1
,2
]}

Quispe, Ruth L. ^{[3
]}

Guzman, Frank ^{[4
]}

Margis, Rogerio ^{[5
]}

Nazari, Evelise M. ^{[1
]}

Muller, Yara M. R. ^{[1
]}

机构：

[1] Univ Fed Santa Catarina, Dept Biol Celular Embriol & Genet, BR-88040900 Florianopolis, SC, Brazil

[2] Ctr Univ Catolica Santa Catarina, BR-89203005 Joinville, SC, Brazil

[3] Univ Fed Santa Catarina, Programa Posgrad Neurociencias, Campus Univ, BR-88040900 Florianopolis, SC, Brazil

[4] Univ Fed Rio Grande do Sul, PPGBCM, Ctr Biotecnol, BR-91501970 Porto Alegre, RS, Brazil

[5] Univ Fed Rio Grande do Sul, Dept Biofis, BR-91501970 Porto Alegre, RS, Brazil

来源：

GENE | 2017年 / 598卷

关键词：

Embryo; Expression stability; Normalization; Gene expression; Prawn; Crustacea; TRANSCRIPTION-PCR DATA; REAL-TIME PCR; MOLECULAR CHARACTERIZATION; GENOMIC ORGANIZATION; SELECTION; ROSENBERGII; PRAWN; VALIDATION; ACCURATE; QUANTIFICATION;

D O I：

10.1016/j.gene.2016.11.001

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

RT-qPCR is a sensitive and highly efficient technique that is widely used in gene expression analysis and to provide insight into the molecular mechanisms underlying embryonic development. The freshwater prawn, Macrobrachium olfersii is an emerging model organism, but, the stable reference genes of this species need to be identified and validated for RT-qPCR analysis. Thus, the aim of this study was to evaluate the expression stability of six genes (beta-act, GAPDH, EF-1 alpha, RpL8, RpS6, AK) in embryos and in adult tissues (cerebral ganglia, muscle and hepatopancreas) of M. olfersii. The expression stabilities of these genes were evaluated using geNorm, NormFinder, BestKeeper, Delta Ct method and integrated tool RefFinder. In the general ranking, RpL8 and RpS6 were the most stable genes in embryos, while RpS6 and RpL8 were the most stable in a combined adult tissue analysis. Analysis of the adult tissues revealed that beta-act and AK were the most stable genes in cerebral ganglia, RpL8 and AK in muscle, and RpS6 and beta-act in hepatopancreas. EF-1 alpha and GAPDH were the least stable genes and as normalizer genes in RT-qPCR affected expression of the Distal-less gene during M. olfersii development. This study provides suitable reference genes for RT-qPCR analysis and allows future studies of the gene expression in M. olfersii for understanding the molecular mechanisms of their development. To our knowledge, this is the first published study that identifies and evaluates reference genes for RT-qPCR analysis in M. olfersii and could be useful as basis for evaluations of reference genes in other prawns. (C) 2016 Elsevier B.V. All rights reserved.

引用

页码：97 / 106

页数：10

共 61 条

[61] Actin gene in prawn, Macrobrachium rosenbergii:: Characteristics and differential tissue expression during embryonic development [J].

Zhu, XJ ;

Dai, ZM ;

Liu, J ;

Yang, WJ .

COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 2005, 140 (04) :599-605

← 1 2 3 4 5 6 7 →