摘要
β-actin mRNA和18S rRNA因其在相同试验条件下的稳定表达，而被广泛用作内标基因来确定目的基因的表达量。试验分别使用这2种内标基因，用相对定量RT—PCR的方法对肉鸡（0-58d）十二指肠钠葡萄糖共转运载体1（Sodium glucose cotransporter 1，SGLT1）的发育规律进行了分析研究，比较不同内标基因对SGLT1 mRNA表达水平检测结果的影响，以确定两者在一致试验条件下表达的相似度。结果表明，分别以β-ctin mRNA和18S rRNA作为内标，SGLT1 mRNA表达发育规律完全一致，均表现为2～30d不断升高，44d下降，58d回升；2d和44d的SGLT1 mRNA丰度显著低于16d（P〈0．05）和30d（P〈0．05），58d的SGLT1 mRNA表达水平显著低于30d（P〈0．05）。提示这2种管家基因均可以作为内标用于肉鸡肠道转运载体基因mRNA发育规律的相对定量研究。
Beta-actin mRNA and 18S rRNA are widely employed as internal control genes, with the assumption that they are expressed constitutively to a similar degree under same experimental conditions. The objective of this study was to investigate the SGLT1 （ Sodium glucose cotransporter 1 ） mRNA ontogenetic expression in duodenum of broiler chicken （0 - 58 d） by a semi-quantitative RT-PCR, using beta-actin mRNA and 18S rRNA as internal standard respectively. In this study, we tested this assumption by assessment of the transcription of these two genes in chicken intestinal tissues using a semi-quantitative RT-PCR. The results indicate showed that the SGLT1 mRNA ontogenetic expression were consistent whether beta-actin mRNA or 18S rRNA was acted as internal control. The SGLT1 mRNA abundance in duodenum of chicken increased from day 2 to day 30, then started to decline from day 44, and increased again from day 58. The SGLT1 mRNA expression level on day 2 and day 44 were lower significantly than that on day 16（P 〈 0.05） and day 30 （P 〈 0.05） respectively. And the SGLT1 mRNA abundance on day 58 was lower significantly than that on day 30 （P 〈 0.05）. It suggested that both beta-actin mRNA and 18S rRNA could be used as internal standard for semi-quanti- tative analysis of transport carrier gene mRNA in intestinal ontogenesis of broiler chicken.
出处
《华北农学报》
CSCD
北大核心
2006年第3期112-116,共5页
Acta Agriculturae Boreali-Sinica
基金
国家重点基础研究发展计划(973)项目(2004CB117501)
广东省自然科学基金项目(05300836)
