华北农学报Issue(1)：154-157,4.DOI:10.7668/hbnxb.2015.01.025
瘤胃脂肪分解菌实时荧光定量PCR方法的构建
Development of a Real-time Quantitative PCR for Anaerovibrio lipolytica in Rumen
摘要
Abstract
To quantify the population of Anaerovibrio lipolytica in rumen,the standard plasmids and curves was constructed by using quantitative Real-time PCR.Total DNA of rumen microbial was isolated from rumen liquid in the Inner Mongolia sheep ,linked with pMD19-T Vector to construct recombinant plasmid and transformed into Esch-erichia coli.White colonies including target plasmids were selected by ampicillin screening ,and the specificity was i-dentified by PCR and DNA sequencing .Plasmid DNA was extracted and the concentration of DNA was measured . The concentration of gradient plasmid DNA was used to construct standard curve for Anaerovibrio lipolytica.Similari-ty of PCR products and purpose gene is more than 99%,correlation coefficient of the standard curve is close to 1 , and the melting peak showed single curve .It was shown that the standard plasmids and curves were successfully constructed and could be applied to quantify the population of unknown Anaerovibrio lipolytica in rumen samples u-sing Real-time PCR.关键词
瘤胃脂肪分解菌/实时荧光定量PCR/标准品质粒/标准曲线Key words
Ruminal Anaerovibrio lipolytica/Real-time quantitative PCR/Standard plasmid/Standard curve分类
农业科技引用本文复制引用
范文斌,刘永斌,李长青..瘤胃脂肪分解菌实时荧光定量PCR方法的构建[J].华北农学报,2015,(1):154-157,4.基金项目
内蒙古自然科学基金项目（2012MS0417）；国家自然科学基金项目 （）
