中国兽医科学2011，Vol.41Issue(3)：273-279,7.
表达新城疫病毒F基因的重组鸡痘病毒实时荧光定量PCR检测方法的建立
Development of real-time PCR method for detection of recombinant fowlpox virus coexpressing F gene of Newcastle disease virus
摘要
Abstract
In order to quantitatively analyze the expression of F gene of Newcastle disease virus(NDV) in recombinant fowlpox virus rFPV-F-VP0,the method with good reproducibility, high specificity and sensitivity standard was established after the real-time PCR reaction conditions were optimized by fluorescent dying methods of SYBR Green. The expression level of NDV F gene in the recombinant fowlpox virus rFPV-F-VP0 was quantitatively analyzed in passages 1,5,10,15 and 20, respectively. The results demonstrated that the standard curve established with the recombinant plasmid showed a fine linear relationship between threshold cycle and template concentration with good reproducibility and specificity. In the linear range of dilution,the dissociation curve was specific and the corresponding C, value increased accordingly with the reduction in the amount of the template. The correlation coefficient and the amplification efficiency was 0.99 ＜r2＜1 and 0.8 ＜E＜1. 2,respectively. This technique accurately quantified the level of exogenous gene expression in the recombinant fowlpox virus,and provided a theoretical basis to clarify the expression mechanism of the recombinant fowlpox virus.关键词
新城疫病毒/F基因/SYBR Green/实时荧光定量聚合酶链反应/鸡痘病毒/基因表达分类
农业科技引用本文复制引用
任静强,鲁会军,金宁一,朱战波,贾鹏,赵权,袭莹,刘存霞,龙川,杜寿文,岳云强..表达新城疫病毒F基因的重组鸡痘病毒实时荧光定量PCR检测方法的建立[J].中国兽医科学,2011,41(3):273-279,7.基金项目
国家高技术研究发展计划(863)项目(2006AA10A205) （863）
