南京农业大学学报2013，Vol.36Issue(1)：92-96,5.DOI:10.7685/j.issn.1000-2030.2013.01.016
实时荧光定量PCR检测猪源Mx1方法的建立及应用
Establishment and application of the SYBR Green Ⅰ real-time PCR assay for detection of porcine antiviral protein Mx1 gene
摘要
Abstract
the basis of the nucleotide sequences of porcine Mxl gene(poMxl)available in GenBank,a pair of primers was designed and synthesized to amplify a specific 115 bp nucleotide fragment from Mxl full-length gene. Then, the SYBR Green I real-time PCR assay was established based on 10-fold dilution recombinant plasmid as the amplification template after it was cloned and sequenced. The results showed that the standard curve regression equation was Y= -2. 986 3 lg X+38. 239 3(R2 =0. 998 5) ,and that the standard curve was of high linearity, specificity, sensitivity and reproducibility. The production of poMxl mRNA from virus-infected cells was quantitatively analyzed after swine kidney cells(PK-15)was stimulated by classical swine fever virus(HCLV strain). The data showed the production of poMxl mRNA was much more after 4 h(P<0. 01) ,and then slowly reduced as time grew. So the results showed that this PCR assay could be applied to detect the expression level of poMxl mRNA.关键词
猪源Mx1/实时荧光定量PCR/标准曲线Key words
porcine Mxl gene(poMxl) /real-time PCR quantitative assay/standard curve分类
农业科技引用本文复制引用
张小敏,周斌,何丹妮,庞然,赵津,陈溥言..实时荧光定量PCR检测猪源Mx1方法的建立及应用[J].南京农业大学学报,2013,36(1):92-96,5.基金项目
国家自然科学基金青年基金项目(31001062) （31001062）
江苏高校优势学科建设工程资助项目 （）
