牛多杀性巴氏杆菌外膜蛋白H的表达与纯化

Abstract

Abstract: In this assay,cloning, expression and identification of outer membrane protein H (OmpH) of Pasteurella multocida were conducted to provide the foundation for study of immune effects of OmpH. OmpH gene was amplified from the genomic DNA of P.multocida CVCC448 by using PCR. After being cloned into pMD18-T, OmpH-pMD18-T was sequenced to confirm its identity, and then cloned into expression vector pET-28a. The constructed recombinant plasmid OmpH-pET28a was transformed into E.coli BL21 and induced with IPTG, and the expressed product was identified by SDS-PAGE and Western blotting. Sequence analysis showed that the full coding length of OmpH was 978 bp, which could encode 326 amino acids, expressed recombinant protein was 37 ku. Western blotting result showed that recombinant OmpH could react with sera of rats immunized with P.multocida, which showed that the recombinant protein had good immunogenicities. OmpH-pET28a was constructed successfully, and fusion protein was expressed in E.coli BL21,which laid the foundation for the study of immune effects of OmpH.

Key words: Pasteurella multocida; outer membrane protein H; clone; expression; identification

CLC Number:

S855.1

HU Xu, LIANG Hong-ru, JIANG Dong-jun, ZHAO Da, GAO Jia-bin, CHEN Wei-hong, YIN Hui, QIAO Bo, ZHU Zhan-bo. Expression and Purification of Outer Membrane Protein H of P. multocida[J]. , 2013, 40(11): 38-41.

References

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Expression and Purification of Outer Membrane Protein H of P. multocida

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