【作者】 胡海； 钱琨； 张丹阳； 石亚运； 张杰； 邵红霞； 金文杰； 秦爱建；

【机构】 扬州大学江苏省动物预防医学重点实验室； 扬州大学教育部禽类预防医学重点实验室； 江苏省动物重要疫病与人兽共患病防控协同创新中心；

【摘要】 目的建立一种检测禽白血病病毒衣壳蛋白p27(ALV-p27)基因绝对荧光定量PCR的方法。方法设计ALVp27特异性荧光定量PCR引物扩增基因并制备质粒标准品,同时建立检测ALV-p27的绝对荧光定量PCR方法。利用建立的绝对定量PCR方法检测病毒感染一日龄雏鸡不同组织脏器中p27基因的分布和表达情况。结果成功建立了检测ALV-p27基因绝对荧光定量PCR的方法。一日龄雏鸡组织脏器中,ALV-p27基因在脑组织中表达水平最高,而在胸腺组织中表达水平最低。结论本研究建立的检测ALV-p27基因绝对定量PCR的方法为进一步研究ALV-p27的生物学功能提供了一种技术手段。更多还原

【Abstract】 Objective To establish a quantitative real-time PCR(qPCR) method for detection of avian leukosis virus caspid protein p27(ALV-p27).Method A pair of specific p27 primers was designed according to the p27 sequence published in genBank.p27 gene was amplified and constructed a standard plasmid DNA,then a standard curve was obtained in the qPCR assay.Distribution of ALV-p27 gene and copies in the different tissues of virus infected one day-old chicken were characterized by qPCR assay.Result An absolute qPCR assay for detection of ALV-p27 was developed.The results indicated that among the different tissues of virus infected one day-old chicken,the p27 gene copy number in Brain was highest,while the thymus showed the lowest gene copy numbers.Conclusion The developed qPCR assay provides a good technology for the further bio-functional study of ALV-p27.更多还原