Transcriptome-based evaluation and validation of suitable housekeeping gene for quantification real-time PCR under specific experiment condition in teleost fishes

	Transcriptome-based evaluation and validation of suitable housekeeping gene for quantification real-time PCR under specific experiment condition in teleost fishes
	Li, Yunkun; Han, Jiabei; Wu, Jiayu; Li, Dong; Yang, Xixi; Huang, Anqi; Bu, Guixian; Meng, Fengyan; Kong, Fanli; Cao, Xiaohan; Han, Xingfa; Pan, Xiaofu; Yang, Shiyong; Zeng, Xianyin; Du, Xiaogang
	2020
发表期刊	FISH & SHELLFISH IMMUNOLOGY
ISSN	1050-4648
卷号	98 页码:218-223
摘要	Quantification real-time PCR (qRT-PCR) is a common method in analysis of gene expression, but the stable reference genes for the normalization analysis have not been appreciated before identifying expression pattern of genes in teleost fishes. In this study, we selected eight candidate reference genes (18S, Actin, EF-1 alpha, 40S, B2M, TUBA, UBCE and GAPDH) basing on transcriptome analysis and the traditional housekeeping genes, and analyzed the stability of the reference genes in spleen, head kidney and head kidney leukocytes (HKL) after pathogen challenge in Schizothorax prenanti (S. prenanti). Three common programs (geNorm, NormFinder and Bestkeeper) were used to evaluate the stability of the candidate reference genes. Two reference genes, Actin and EF-1 alpha presented higher stability, while 18S and GAPDH were the lower stable genes, both in in vitro and in vivo. An important immune gene, toll-like receptor 22a (TLR22a), was selected to validate the stability of the proposed reference genes (Actin and EF-1 alpha) across different experiment treatments. The results reveal that Actin and EF-1 alpha are quite suitable reference genes for the normalization analysis. Otherwise, using the most stable gene Actin to validate the reliable of transcriptome data showed the high correlation between the fold change of transcriptome data and qRT-PCR data. In conclusion, our study not only acquired the suitable reference gene for the qRT-PCR assay under specific experiment condition, but also provided a comprehensive method to evaluate and validate the reference gene based on transcriptome analysis in teleost fishes.
收录类别	sci
语种	英语
文献类型	期刊论文
条目标识符	http://ir.kiz.ac.cn/handle/152453/12770
专题	科研部门_系统进化与生物地理学（杨君兴）
推荐引用方式 GB/T 7714	Li, Yunkun,Han, Jiabei,Wu, Jiayu,et al. Transcriptome-based evaluation and validation of suitable housekeeping gene for quantification real-time PCR under specific experiment condition in teleost fishes[J]. FISH & SHELLFISH IMMUNOLOGY,2020,98:218-223.
APA	Li, Yunkun.,Han, Jiabei.,Wu, Jiayu.,Li, Dong.,Yang, Xixi.,...&Du, Xiaogang.(2020).Transcriptome-based evaluation and validation of suitable housekeeping gene for quantification real-time PCR under specific experiment condition in teleost fishes.FISH & SHELLFISH IMMUNOLOGY,98,218-223.
MLA	Li, Yunkun,et al."Transcriptome-based evaluation and validation of suitable housekeeping gene for quantification real-time PCR under specific experiment condition in teleost fishes".FISH & SHELLFISH IMMUNOLOGY 98(2020):218-223.

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