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马子宫内膜、睾丸和孕体组织中假定参考基因的表达稳定性

Expression stability of putative reference genes in equine endometrial, testicular, and conceptus tissues.

作者信息

Klein Claudia, Rutllant Josep, Troedsson Mats Ht

机构信息

University of Kentucky, Department of Veterinary Science, 108 Gluck Equine Research Center, Lexington, KY, 40546, USA.

出版信息

BMC Res Notes. 2011 Apr 12;4:120. doi: 10.1186/1756-0500-4-120.

DOI:10.1186/1756-0500-4-120
PMID:21486450
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3083352/
Abstract

BACKGROUND

Quantitative RT-PCR data are commonly normalized using a reference gene. A reference gene is a transcript which expression does not differ in the tissue of interest independent of the experimental condition. The objective of this study was to evaluate the stability of mRNA expression levels of putative reference genes in three different types of equine tissue, endometrial, testicular, and conceptus tissue.

FINDINGS

The expression stability of four (uterine tissue) and six (testicular and conceptus tissue) was assessed using descriptive data analysis and the software programs Normfinder and geNorm. In uterine samples, 18S showed the largest degree of variation in expression while GAPDH, B2M, and ACTB were stably expressed. B2M and GAPDH were identified as the most stably expressed genes in testicular samples, while 18S showed some extent of regulation between samples. Conceptus tissue overall was characterized by very low variability of the transcripts analyzed with GAPDH, YWHZ, and 18S being the most stably expressed genes.

CONCLUSIONS

In equine endometrium, GAPDH, B2M, and ACTB transcript levels are equally stable, while 18S is less stably expressed. In testes and associated structures, B2M and GAPDH are the transcripts showing the least amount of variation, while in conceptus tissue GAPDH, YWHZ, and 18S were identified as the most suitable reference genes. Overall, transcripts analyzed in conceptus tissue were characterized by less variation than transcripts analyzed in uterine and testicular tissue.

摘要

背景

定量逆转录聚合酶链反应（qRT-PCR）数据通常使用参考基因进行标准化。参考基因是一种转录本，其在感兴趣的组织中的表达不受实验条件的影响而有所不同。本研究的目的是评估三种不同类型马组织（子宫内膜、睾丸和孕体组织）中假定参考基因mRNA表达水平的稳定性。

研究结果

使用描述性数据分析以及Normfinder和geNorm软件程序评估了四种（子宫组织）和六种（睾丸和孕体组织）基因的表达稳定性。在子宫样本中，18S的表达变化程度最大，而甘油醛-3-磷酸脱氢酶（GAPDH）、β2微球蛋白（B2M）和β-肌动蛋白（ACTB）表达稳定。B2M和GAPDH被确定为睾丸样本中表达最稳定的基因，而18S在样本间表现出一定程度的调控。孕体组织总体上的特点是所分析转录本的变异性非常低，GAPDH、酪氨酸3-单加氧酶/色氨酸5-单加氧酶激活蛋白ζ（YWHZ）和18S是表达最稳定的基因。

结论

在马的子宫内膜中，GAPDH、B2M和ACTB转录水平同样稳定，而18S表达较不稳定。在睾丸及相关结构中，B2M和GAPDH是变异程度最小的转录本，而在孕体组织中，GAPDH、YWHZ和18S被确定为最合适的参考基因。总体而言，孕体组织中分析的转录本的变异程度低于子宫和睾丸组织中分析的转录本。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8392/3083352/bbf00c6197a0/1756-0500-4-120-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8392/3083352/d83888745959/1756-0500-4-120-1.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8392/3083352/ce130cc1cbf5/1756-0500-4-120-7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8392/3083352/bbf00c6197a0/1756-0500-4-120-8.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8392/3083352/d83888745959/1756-0500-4-120-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8392/3083352/65c98af1b3ef/1756-0500-4-120-2.jpg
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