猪肺炎支原体P65基因原核表达及免疫原性分析

doi:10.11843/j.issn.0366-6964.2014.02.014

Abstract

Abstract:

Protein P65，the immunodominance protein of Mycoplasma hyopneumoniae (Mhp)，and shows no cross reaction with other Mycoplasmas，is usually used as the target protein or gene for Mhp detection.In this study，the front 922 bp fragment containing three rare codons in P65 gene was obtained by gene synthesis，and the other fragment of P65 gene was amplified by PCR from Mhp 168 strain，then the mutated P65 gene was obtained through the Splicing by overhang extension (SOE-PCR) with the surplus segment connection，finally the recombinant plasmid pET-32a(+)/P65 was constructed by inserting the mutated P65 gene into expression vector pET-32a(+).Under IPTG induction，E.coli BL21(DE3)/pET-32a(+)/P65 expressed the recombinant P65 protein with 87 kD.The result of Western blot analysis showed that the recombinant protein has satisfactory immunoreactivity.High level of antibody was obtained in BALB/c mice after inoculating the recombinant P65 protein，which indicated the recombinant P65 protein has a good immunogenicity.We had successfully obtained the recombinant P65 protein of Mhp by expressing in the E.coli，it provide the support for Mhp detection and subunit vaccine，and also offers the help for the further study of the pathogenic mechanism of this disease.

CLC Number:

S852.62

LIU Mao-jun，ZHANG Yue，DU Gai-mei，BAI Yun，WANG Hai-yan，JIANG Ping，SHAO Guo-qing. Prokaryotic Expression and Immunogenicity Identification of P65 from Mycoplasma hyopneumoniae[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(2): 262-267.

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Prokaryotic Expression and Immunogenicity Identification of P65 from Mycoplasma hyopneumoniae

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