ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2012, Vol. 43 ›› Issue (8): 1324-1329.

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Expression of P216 Gene Fragment from Mycoplasma hyopneumoniae and Research on Adhesion Activity

DU Haixia 1,2，LIU Maojun 1,2，FENG Zhixin 1，XIONG Qiyan 1， BAI Fangfang 1，WANG Haiyan 1，SHAO Guoqing 1*   

  1. 1. Key Laboratory of Veterinary Biological Engineering and Technology Ministry of Agriculture National Center for Engineering Research of Veterinary Bioproducts, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China; 2. College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
  • Received:2012-01-06 Revised:1900-01-01 Online:2012-08-24 Published:2012-08-24
  • Contact: SHAO Guoqing

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Abstract: This experiment was conducted to study the adhesion activity of P216 protein and establish the model of adhesion protein of Mycoplasma hyopneumoniae (Mhp). According to analysis, the fragment of P216 gene with hydrophilicity, antigenicity and good adhesion was chose. P216 gene fragment was amplified by PCR from Mhp NJ strain and inserted into expression vector pET32a(+), and the recombinant plasmid pET32a(+)/P216 was constructed. After IPTG induction, the immunological and adhesion activity of the recombined protein was detected by Western blot and indirect immunofluorescence assay. The results showed that, the PCR product of the target gene was l 636 bp, the molecular weight of recombinant protein was 80.1 kDa by SDSPAGE, and Western blot results showed that recombinant protein had satisfactory immunogenicity. Indirect immunofluorescence assay showed that the recombinant protein could produce occupied inhibition to the Mhp adhering with SJPL cells. These results indicated that the P216 protein had good adhesion activity, and could adhere SJPL cells. It provides new ideas for research of the other adhesions from Mycoplasma hyopneumoniae.

Key words: Mycoplasma hyopneumoniae, P216 gene fragment, adhesion