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根据GenBank中公布的原鸡beta-连环蛋白(β-catenin)参考序列设计特异性引物扩增得到β-catenin基因,并克隆到真核表达载体pcDNA3.1中作为荧光定量PCR标准品;同时利用软件设计家禽β-catenin的荧光定量PCR引物,建立检测β-catenin的绝对荧光定量PCR方法。并利用该方法检测病毒感染组与非感染组1日龄雏鸡不同组织中β-catenin基因的分布和表达情况。结果显示,在感染和非感染病毒的1日龄雏鸡中,β-catenin基因在脑组织中表达水平最高,而在脾脏和法氏囊中表达水平较低。结果表明,成功建立了检测家禽β-catenin基因的绝对定量PCR方法,并检测了β-catenin基因在1日龄雏鸡不同脏器中的表达情况,为进一步研究其生物学功能提供了技术手段。
The β-catenin gene was amplified by designing a specific primer according to the beta-catenin reference sequence published in GenBank and cloned into the eukaryotic expression vector pcDNA3.1 as a fluorescent quantitative PCR standard. At the same time, Software design of poultry β-catenin fluorescence quantitative PCR primers, the establishment of detection of β-catenin absolute fluorescence quantitative PCR method. The method was used to detect the distribution and expression of β-catenin in different tissues of 1-day-old chickens infected with virus and non-infected chickens. The results showed that in the 1-day-old chickens infected and not infected with virus, β-catenin gene in brain tissue expression was the highest level, while in the spleen and bursa of lower expression levels. The results showed that the absolute quantitative PCR method of β-catenin gene detection in poultry was successfully established and the expression of β-catenin gene in different organs of 1-day-old chickens was detected, which provided a technical means for further study of its biological function.