猫子宫内膜中内参基因的验证

Validation of reference genes in the feline endometrium.

作者信息

Jursza Ewelina, Skarzynski Dariusz J, Siemieniuch Marta J

机构信息

Department of Reproductive Immunology and Pathology, Institute of Animal Reproduction and Food Research of the Polish Academy of Sciences, Tuwima 10, 10-748 Olsztyn, Poland.

出版信息

Reprod Biol. 2014 Dec;14(4):302-6. doi: 10.1016/j.repbio.2014.04.003. Epub 2014 Apr 30.

DOI:10.1016/j.repbio.2014.04.003

PMID:25454498

Abstract

The aim of the study was to find the most stable reference genes from: ACTB, GAPDH, RPL30, CYC, RPL17, RPS7 and YWHAZ in the feline endometrium. Three free software packages, geNorm, NormFinder and BestKeeper were used. In geNorm analysis, the most stable gene was RPS7 (at a primer concentration 1000 nM) or YWHAZ (500 and 250 nM). According to NormFinder and BestKeeper, ACTB (at all examined primer concentrations) followed by RPS7 and CYC were the most stable genes. Based on geNorm results at least two genes from among RPS7, RPL30, ACTB or YWHAZ should be chosen for Real Time-PCR result normalization.

摘要

本研究的目的是在猫子宫内膜中从以下基因

β-肌动蛋白（ACTB）、甘油醛-3-磷酸脱氢酶（GAPDH）、核糖体蛋白L30（RPL30）、细胞色素C（CYC）、核糖体蛋白L17（RPL17）、核糖体蛋白S7（RPS7）和14-3-3蛋白ζ/δ（YWHAZ）中找到最稳定的内参基因。使用了三个免费软件包，即geNorm、NormFinder和BestKeeper。在geNorm分析中，最稳定的基因是RPS7（引物浓度为1000 nM时）或YWHAZ（500 nM和250 nM时）。根据NormFinder和BestKeeper的结果，ACTB（在所有检测的引物浓度下）其次是RPS7和CYC是最稳定的基因。基于geNorm的结果，应从RPS7、RPL30、ACTB或YWHAZ中选择至少两个基因用于实时定量PCR结果的标准化。