Validation of Reference Genes for Real-Time PCR of Reproductive System in the Black Tiger Shrimp

被引：34

作者：

Leelatanawit, Rungnapa ^{[1
]}

Klanchui, Amornpan ^{[1
]}

Uawisetwathana, Umaporn ^{[1
]}

Karoonuthaisiri, Nitsara ^{[1
]}

机构：

[1] Natl Sci & Technol Dev Agcy, Natl Ctr Genet Engn & Biotechnol, Microarray Lab, Klongluang, Pathumthani, Thailand

来源：

PLOS ONE | 2012年 / 7卷 / 12期

关键词：

MESSENGER-RNA EXPRESSION; HOUSEKEEPING GENES; BETA-ACTIN; INTERNAL CONTROL; RT-PCR; GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE; MOLECULAR-CLONING; ATLANTIC SALMON; DIFFERENTIAL EXPRESSION; TESTICULAR DEVELOPMENT;

D O I：

10.1371/journal.pone.0052677

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Gene expression of reproductive system of the black tiger shrimp (Peneaus monodon) has been widely studied to address poor maturation problem in captivity. However, a systematic evaluation of reference genes in quantitative real-time PCR (qPCR) for P. monodon reproductive organs is lacking. In this study, the stability of four potential reference genes (18s rRNA, GAPDH, beta-actin, and EF1-alpha) was examined in the reproductive tissues in various conditions using bioinformatic tools: NormFinder and geNorm. For NormFinder, EF1-alpha and GAPDH ranked first and second as the most stable genes in testis groups whereas GAPDH and EF1-alpha were for ovaries from wild-caught broodstock and domesticated groups. EF1-alpha and beta-actin ranked first and second for the eyestalk ablated ovaries. For geNorm, EF1-alpha and GAPDH had the best stability in all testis and ovaries from domesticated groups whereas EF1-alpha and beta-actin were the best for ovaries from wild-caught and eyestalk ablated groups. Moreover, the expression levels of two well-known reproductive genes, Dmc1 and Vitellogenin, were used to validate these reference genes. When normalized to EF1-alpha, the expected expression patterns were obtained in all cases. Therefore, this work suggests that EF1-alpha is more versatile as reference genes in qPCR analysis for reproductive system in P. monodon.

引用

页数：10

共 67 条

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