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 Escherichia coli.White colonies including target plasmids were selected by ampicillin screening, and the specificity was identified 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.Similarity 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 using Real-time PCR.

Key words: Ruminal Anaerovibrio lipolytica, Real-time quantitative PCR, Standard plasmid, Standard curve

CLC Number: 

• S826.8