禽白血病病毒衣壳蛋白p27基因绝对定量PCR方法的建立及应用

Establishment and Application of an Absolute Quantification Real-time PCR Assay for Detection of Capsid Protein p27 Gene of Avian Leukosis Virus

研究旨在建立一种检测禽白血病病毒(ALV)衣壳蛋白p27(ALV-p27)基因实时荧光定量PCR的方法。通过设计ALV-p27特异性的荧光定量PCR引物扩增基因并制备质粒标准品,同时建立检测病毒衣壳蛋白p27基因的实时荧光定量PCR方法,并利用该方法检测6日龄胚胎感染,1日龄出壳雏鸡不同组织中p27基因的分布和表达情况。结果表明,ALV-p27基因在1日龄雏鸡脑组织中表达水平最高,而在胸腺组织中表达水平最低。研究建立的检测ALV-p27基因绝对定量PCR方法为进一步研究其生物学功能提供了一种技术手段。

The purpose of this study was to establish an absolute quantification real-time PCR method for detection of avian leukosis virus （ALV） caspid protein p27 （ALV-p27）. By using the designed specific p27 primer pairs,p27 gene was amplified and used to make standard plasmid DNA which was used to create the standard curve in the absolute quantification real-time PCR assay. And ALV-p27 gene distribution and copies in the different tissues of one-day-age chicken injected by ALV （JS09GY03） at 6th day of embryonation via yolk sac were characterized by the real-time PCR assay. The results indicated that among the different tissues of one-day-age infected chicken,copy number of p27 gene in brain was the highest, while the lowest number was in the thymus. The established absolute quantification real-time PCR assay could provide an aviable method for the further bio-functional study of ALV-p27.