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KMID : 0545120040140061343
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Journal of Microbiology and Biotechnology
2004 Volume.14 No. 6 p.1343 ~ p.1349
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Molecular Cloning and Characterization of the Gene for Outer Membrane Protein H in a Pasteurella multocida (D:4) Isolate from Pigs with Atrophic Rhinitis Symptoms in Korea
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Lee JM
Kang SY/Park SI/Woo HJ/Kwon MS
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Abstract
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A native strain of Pasteurella multocida was isolated from pigs suffering from severe atrophic rhinitis at domestic farms in Gyeonggi Province Korea and was identified as capsular serogroup “D” and somatic serotype “4” by disc diffusion decapsulation and gel diffusion precipitation tests respectively. The P. multocida (D:4) induced atrophic rhinitis in healthy pigs by the secondary infection. The gene for outer membrane protein H (ompH) of P. multocida (D:4) was cloned in Escherichia coli DH5α by PCR. The open reading frame of the ompH was composed of 1023 bp possibly encoding a protein with 341 amino acid residues containing a signal peptide of 20 amino acids at N-terminus and the gene product with molecular mass of ca. 38 kDa was identified by SDS-PAGE. Hydropathy profiles indicated that there are two variable domains in the OmpH. To express the ompH in E. coli the gene was manipulated in various ways. Expression of the truncated as well as full-length forms of the recombinant OmpH was fatal to the host E. coli BL21 (DE3). However the truncated OmpH fused with GST was consecutively expressed in E. coli DH5α. A large quantity of the fused polypeptide was purified through GST-affinity chromatography.
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