Selection and Validation of Reference Genes for RT-PCR Expression Analysis of Candidate Genes Involved in Morphine-Induced Conditioned Place Preference Mice

The reliability of qRT-PCR results depend on the stability of reference genes used for normalization, suggesting the necessity of identification of reference genes before gene expression analysis. Morels are edible mushrooms well-known across the world and highly prized by many culinary kitchens. Here, several candidate genes were selected and designed according to the Morchella importuna transcriptome data. The stability of the candidate genes was evaluated with geNorm and NormFinder under three different experimental conditions, and several genes with excellent stability were selected. The extensive adaptability of the selected genes was tested in ten Morchella species. Results from the three experimental conditions revealed that ACT1 and INTF7 were the most prominent genes in Morchella, CYC3 was the most stable gene in different development stages, INTF4/AEF3 were the top-ranked genes across carbon sources, while INTF3/CYC3 pair showed the robust stability for temperature stress treatment. We suggest using ACT1, AEF3, CYC3, INTF3, INTF4 and INTF7 as reference genes for gene expression analysis studies for any of the 10 Morchella strains tested in this study. The stability and practicality of the gene, vacuolar protein sorting (INTF3), vacuolar ATP synthase (INTF4) and14-3-3 protein (INTF7) involving the basic biological processes were validated for the first time as the candidate reference genes for quantitative PCR. Furthermore, the stability of the reference genes was found to vary under the three different experimental conditions, indicating the importance of identifying specific reference genes for particular conditions.

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Identification of suitable reference genes for gene expression analysis of pork meat quality and analysis of candidate genes associated with the trait drip loss

Meat Science ◽

10.1016/j.meatsci.2010.05.030 ◽

2010 ◽

Vol 86 (2) ◽

pp. 436-439 ◽

Cited By ~ 17

Author(s):

Jean McBryan ◽

Ruth M. Hamill ◽

Grace Davey ◽

Peadar Lawlor ◽

Anne Maria Mullen

Keyword(s):

Gene Expression ◽

Candidate Genes ◽

Meat Quality ◽

Expression Analysis ◽

Reference Genes ◽

Gene Expression Analysis ◽

Drip Loss ◽

Pork Meat ◽

Suitable Reference

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Validation of reference genes for quantitative expression analysis by real-time RT-PCR in Saccharomyces cerevisiae

BMC Molecular Biology ◽

10.1186/1471-2199-10-99 ◽

2009 ◽

Vol 10 (1) ◽

pp. 99 ◽

Cited By ~ 290

Author(s):

Marie-Ange Teste ◽

Manon Duquenne ◽

Jean M François ◽

Jean-Luc Parrou

Keyword(s):

Saccharomyces Cerevisiae ◽

Real Time ◽

Expression Analysis ◽

Reference Genes ◽

Rt Pcr ◽

Quantitative Expression

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Expression Analysis of the Prolific Candidate Genes, BMPR1B, BMP15, and GDF9 in Small Tail Han Ewes with Three Fecundity (FecB Gene) Genotypes

Animals ◽

10.3390/ani8100166 ◽

2018 ◽

Vol 8 (10) ◽

pp. 166 ◽

Cited By ~ 11

Author(s):

Jishun Tang ◽

Wenping Hu ◽

Ran Di ◽

Qiuyue Liu ◽

Xiangyu Wang ◽

...

Keyword(s):

Gene Expression ◽

Candidate Genes ◽

Expression Pattern ◽

Expression Analysis ◽

Regulatory Mechanism ◽

Follicular Development ◽

Tissue Expression ◽

Rt Pcr ◽

Tissue Expression Pattern ◽

Systematic Analysis

The expression characteristics of the prolific candidate genes, BMPR1B, BMP15, and GDF9, in the major visceral organs and hypothalamic–pituitary–gonadal (HPG) axis tissues of three FecB genotypes (FecB BB, FecB B+, and FecB ++) were explored in STH ewes using RT-PCR and qPCR. The results were as follows, BMPR1B was expressed in all FecB BB genotype (Han BB) tissues, and GDF9 was expressed in all selected tissues, but BMP15 was specifically expressed in the ovaries. Further study of ovarian expression indicated that there was no difference in BMPR1B expression between genotypes, but the FecB B+ genotype (Han B+) had greater expression of GDF9 and BMP15 than Han BB and FecB ++ genotype (Han ++) (p < 0.05, p < 0.01). BMP15 expression was lower in the ovaries of Han BB than in Han ++ sheep, but the reverse was shown for GDF9. The gene expression in non-ovarian tissues was also different between genotypes. Therefore, we consider that the three genes have an important function in ovine follicular development and maturation. This is the first systematic analysis of the tissue expression pattern of BMPR1B, BMP15, and GDF9 genes in STH sheep of the three FecB genotypes. These results contribute to the understanding of the molecular regulatory mechanism for ovine reproduction.

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