鸡p53 mRNA实时荧光定量PCR检测方法的建立及应用  

Establishment and Application of SYBR GREENⅠReal-time PCR for Detection of p53 mRNA Expression in Chickens

作　　者：佟美姣[1,2] 高雪丽 郑世民[1] TONG Mei-jiao;GAO Xue-li;ZHENG Shi-min(Northeast Agricultural University,Harbin,Heilongjiang,150030,China;Baoqing Center for Animal Disease Prevention and Control,Shuangyashan,Heilongjiang,155600,China)

机构地区：[1]东北农业大学,黑龙江哈尔滨150030 [2]宝清县动物疫病预防与控制中心,黑龙江省双鸭山155600

出　　处：《动物医学进展》2021年第11期24-28,共5页Progress In Veterinary Medicine

基　　金：国家自然科学基金青年基金(30901062)。

摘　　要：p53肿瘤蛋白由p53基因编码,在机体内有多种作用,例如,修复损伤DNA,调控细胞周期以及细胞凋亡,能够抑制肿瘤的发生。根据已发表的鸡p53 mRNA相关基因序列设计引物,提取鸡免疫器官总RNA,通过转录、纯化、链接等步骤,以提取的重组阳性质粒为标准品进行检测,绘制标准曲线,并对其特异性进行检测,建立了鸡p53 mRNA实时荧光定量检测方法。应用该方法对雏鸡免疫器官的p53 mRNA表达量进行检测,发现传染性法氏囊病病毒(Infectious bursal disease virus,IBDV)感染21日龄SPF雏鸡后,其胸腺与脾脏中p53 mRNA表达第3天显著高于相应对照雏鸡;法氏囊中p53 mRNA表达量在病毒感染后5 d~7 d显著或极显著增高。试验结果表明,此检测方法重复性好,可操作性强,可以用于相关基因的检测。The p53 gene encodes P53 protein that has many functions in the boby,such as,repaires damaged DNA,regulates the cycle of cell and apoptosis,inhibites the occurrence of tumor.According to chicken p53 gene sequences,the pairs of primers were successfully designed.The total of RNA was extracted from the immune organs of chicken.Through a series of steps,such as transcription,purification,linking,etc.,the recombinant positive plasmid was as the standard substance,which was to develop the assay and to draw the standard curve.The real-time fluorescence quantitativ detection method of chicken p53 mRNA was successfully established which was used to detect the expression of p53 mRNA in the immune organs of chicks.The results showed that:after IBDV infected 21 days SPF chickens,the expression of p53 mRNA in thymus and spleen of infected chickens were much higher than of the control chickens in 3 days.The expression of p53 mRNA in bursal of Fabricius of infected chickens were significant and very significant higher than of the control chickens in 5-7 days.The test has good repeatability and strong operability,which provided a reliable basis for further detection work.

关 键 词：P53基因 实时荧光定量PCR 鸡 

分 类 号：S852.2[农业科学—基础兽医学] S858.31[农业科学—兽医学]