Identification of housekeeping genes suitable for gene expression analysis in Jian carp (Cyprinus carpio var. jian)

文献类型： 外文期刊

第一作者： Tang, Yong-kai

作者： Tang, Yong-kai;Yu, Ju-hua;Xu, Pao;Li, Jian-lin;Li, Hong-xia;Tang, Yong-kai;Yu, Ju-hua;Xu, Pao;Ren, Hong-tao

作者机构：

关键词： Quantitative real time RT-PCR;Housekeeping gene;Cloning;Cyprinus carpio var. jian

期刊名称：FISH & SHELLFISH IMMUNOLOGY （ 影响因子：4.581； 五年影响因子：4.851 ）

ISSN： 1050-4648

年卷期： 2012 年 33 卷 4 期

页码：

收录情况： SCI

摘要： Jian carp (Cyprinus carpio var. jian) is an important economic fish species cultured in China. In this report, we performed a systematic analysis to identify an appropriate housekeeping (HK) gene for the study of gene expression in Jian carp. For this purpose, partial DNA sequences of four potential candidate genes (elongation factor 1 alpha (EF-1 alpha), glyceraldehyde-3-phosphate (GAPDH), beta-actin (ACTB), and 18S ribosomal RNA (18S rRNA) were isolated, and their expression levels were studied using RNA extracted from nine tissues (forebrain, hypothalamus, liver, fore-intestine, hind-intestine, ovary, muscle, heart, kidney) in juvenile and adult Jian carp. Gene expression levels were quantified by quantitative real time RT-PCR (qRT-PCR), and expression stability was evaluated by comparing the coefficients of variation (CV) of the Ct values. The results showed that EF-1 alpha was the most suitable HK gene in all tissues of juvenile and adult Jian carp. However, at distinct juvenile and adult developmental stages, there was not a single optimal gene for normalization of expression levels in all tissues. EF-1 alpha was the most stable gene only in forebrain, hypothalamus, liver, heart, and kidney. These results provide data that can be expected to aid gene expression analysis in Jian carp research, but underline the importance of identifying the optimal HK gene for each new experimental paradigm. (C) 2012 Elsevier Ltd. All rights reserved.

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