Genetic diversity of dipeptidyl peptidase IV from anaerobic bacterial cultivation in vitro in dairy cow

Abstract Text:

The objective of this experiment was to reveal the genetic diversity of dipeptidyl peptidase IV (DPP-IV) gene from anaerobic bacterial cultivation in vitro and the influence of monensin to bacteria containing DPP-IV gene. There were two groups: one group substrate was casein (P), the other group substrate was casein with monensin added (M). Each group had three duplicated samples. Anaerobic culture was done after inoculating with rumen fluid. And then anaerobic bacterium was collected after 12 h culture and its DNA was extracted. DPP-IV gene library was established through PCR. Samples were detected by quantitative real-time PCR. Sixty-six sequence of DPP-IV gene were obtained from picking clones. The results of BLASTP analysis showed these sequences matched with Prevotella ruminicola, Prevotella marshii and Paraprevotella xylaniphila. Twenty-three operational taxonomic units (OTU) were found by Mothur software analysis, and OTU9 might be a DPP-IV gene of protein degradation bacteria sensitising to monensin. Quantitative analysis showed that the copy number of DPP-IV gene in P group was significantly higher than that of M group (P<0.01). These results suggested that monensin affected the amount of bacteria containing DPP-IV gene.

Keywords:

anaerobic bacterial cultivation in vitro; dipeptidyl peptidases IV; genetic diversity