Evaluation of reference genes for investigation of gene expression profiles in chorioallantois from Meishan and Yorkshire pigs on gestational days 26 and 50
首发时间:2013-12-30
Abstract:Selection of suitable reference gene is important for accurate measurement of gene expression data obtained using the quantitative real-time PCR (qPCR) method. There has been no systematic investigation of reference genes for porcine chorioallantois which plays important roles in mediating concepts/fetus growth and survival. In this study, six candidate reference genes expressed in pig chorioallantois from Meishan and Yorkshire pigs on gestational days 26 (GD26) and 50 (GD50) were evaluated by qPCR. These genes included TATA box binding protein (TBP), glyceraldehydes-3-phosphate dehydrogenase (GAPDH), ribosomal protein L32 (RPL32), actin-beta (ACTB), ribosomal protein S20 (RPS20) and RNApolymerase polypeptide G (RPG). The expression stability of these genes was investigated by using the GeNorm software program and statistical analyses. The rank order of the expression stability for these genes was (from the most stable to the least stable): TBP, GAPDH, RPL32, ΒACTIN, RPS20 and RPG. Collectively, the results suggest that the TBP and GAPDH could be considered as the suitable reference genes for investigation of gene expression in the pig chorioallantois from the two breeds at different gestational stages.
keywords: Animal Genetics, Breeding and Reproduction Reference gene qPCR Chorioallantois Pig
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梅山和大白猪妊娠早中期胎盘绒毛膜组织基因表达谱内参基因的筛选
摘要:在使用定量PCR(qPCR)对基因表达进行精确定量时,选择合适的内参基因尤其重要。猪的绒毛膜组织对于孕体/胎儿生长乃至存活十分重要,但目前可用于该组织的内参基因还没有系统的研究。本研究选择在梅山猪和大白猪妊娠26天和50天绒毛膜组织均无差异表达的6个内参基因(TBP、GAPDH、RPL32、ACTB、RPS20和RPG),通过qPCR方法,利用GeNorm软件包对他们的表达稳定性进行评估。统计分析结果显示,这6个基因的稳定性依次为TBP> GAPDH>RPL32>ΒACTIN>RPS20 > RPG. 由此可见,TBP和 GAPDH 基因是两个可用于猪妊娠早中期胎盘绒毛膜组织基因表达谱研究的比较合适的内参基因。
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