›› 2017, Vol. 44 ›› Issue (8): 2248-2254.doi: 10.16431/j.cnki.1671-7236.2017.08.005

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Cloning,Expression and Bioinformatics Analysis of Enterococcus faecalis from Northeast Black Bear

QIN Xiao-dong1,2, CHU Xi-tong1, LU Cheng1, SUN Fu-liang1, SUN Xing-zhong1,3   

  1. 1. Agricultural College of Yanbian University, Yanji 133000, China;
    2. Animal Quarantine Station of Dunhua, Dunhua 133700, China;
    3. Animal Disease Prevention and Control Center of Baicheng, Baicheng 137000, China
  • Received:2017-01-06 Online:2017-08-20 Published:2017-08-18

Abstract:

In order to detect Enterococcus faecalis endocarditis antigen (EfaA) of bear that was used to disclose the infected Northeast Black bear immediately, a pair of PCR primers was synthesized by EfaA gene of Enterococcus faecalis that was recorded in GenBank (accession number:U03756.1) and the target fragment was 689 bp. The PCR product was cloned by pMD19-T vector, then was transferred to Escherichia coli DH5α competent cells and the positive clones were filtered. Recombinant plasmid (pMD19-T-EfaA) was extracted. Identification of PCR and digestion, sequencing, the structure prediction of proteins were operated. The results showed that EfaA gene was cloned successfully, and the gene homology with ATCC 29212 was 100.0%. pMD19-T-EfaA was constructed successfully, structure of proteins was predicted. This study provided theoretical basis for diagnostic methods of Enterococcus faecalis and laid basis for prevention and control of Northeast Black bear disease.

Key words: Northeast Black bear; Enterococcus faecalis endocarditis antigen (EfaA); gene cloning; bioinformatics analysis

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