草鱼脂肪组织及脂肪细胞qRT-PCR内参基因的筛选

doi:10.16378/j.cnki.1003-1111.21102

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Abstract To explore the stable expression of the four reference genes in grass carp Ctenopharyngodon idellus adipose tissue and adipocytes, and to select the best stable reference genes, transcription levels of 4 candidate reference genes including 18S rRNA, β-actin, GAPDH and EF1α were investigated in abdominal adipose tissue of grass carp with body weight of (15.0±0.5) g fed diets containing 4%, 8% and 12% of lipid at water temperature of 26.5—27.5 ℃ for 4 weeks when the adipocytes were treated with oleic acid concentrations of 0, 40, 80 and 120 mmol/L by real-time fluorescence quantitative PCR (qRT-PCR), and then the stability of their expression levels were analyzed by software GeNorm, NormFinder and BestKeeper to screen out the reference genes or combinations stably expressed in adipose tissue and adipose cells. The results showed that specific amplification products and ideal amplification efficiency were obtained by all the quantitative PCR primers for candidate reference genes. The descending order of expression stability for candidate reference genes was expressed as 18S rRNA>β-actin>GAPDH>EF1α and 18S rRNA=β-actin>GAPDH>EF1αIn in adipose tissue and adipocytes. The differentiation analysis of standardized factor pairing of GeNorm showed that its V_n/V_{(n +1)}was found to be less than 0.15, that is, with the optimal reference genes number of at least 2 in adipose tissue and adipocytes. The findings indicated that the reference gene stably expressed in adipose tissue and adipocytes of grass carp was 18S rRNA. It was suggested that 18S rRNA and β-actin should be selected as the most suitable reference gene combination in adipose tissue and cells.

Key words： real-time quantitative PCR Ctenopharyngodon idellus screening of reference gene adipose tissue adipocyte expression stability

Received: 15 June 2021

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	HU Zechao
	ZOU Xiaocui
	SUN Jian
	BIAN Chenchen
	JI Hong

Cite this article:

HU Zechao,ZOU Xiaocui,SUN Jian, et al. Identification of Reference Genes in Adipose Tissue and Adipocytes of Grass Carp Ctenopharyngodon idellus by qRT-PCR[J]. Fisheries Science, 2023, 42(4): 547-555.

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https://www.shchkx.com/EN/10.16378/j.cnki.1003-1111.21102 OR https://www.shchkx.com/EN/Y2023/V42/I4/547

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