In order to compare the homology and antigen of Haemophilus parasuis (HPS)4 outer membrane protein P2(omp2), we design a test with specific primers, using PCR amplification of isolates of Haemophilus parasuis from Sic...In order to compare the homology and antigen of Haemophilus parasuis (HPS)4 outer membrane protein P2(omp2), we design a test with specific primers, using PCR amplification of isolates of Haemophilus parasuis from Sichuan Province(HP Sch2010), ompP2 gene will be cloned into the pGM-T vector, and transformed into E. coli DH5α. Identified by PCR and sequencing and analysis, the sequencing results showed that the published 4 HPS SW124 strains omp2 gene (1077bp), compared with the amplified 1086bp purpose fragment(containing omp2 genes), is relatively stable, with the nucleotide homology level 97% and amino acid homology level of 92.5%. The variable regions are mainly concentrated in the three base sequences: 40-65,110-156,180-202.展开更多
Objective: To construct a recombinant plasmid containing the outer membrane protein 2 (Omp2) gene of Chlamydia trachomatis and express Omp2 in E.coli. Methods: The omp2 gene of C. trachomatis serovar D was cloned into...Objective: To construct a recombinant plasmid containing the outer membrane protein 2 (Omp2) gene of Chlamydia trachomatis and express Omp2 in E.coli. Methods: The omp2 gene of C. trachomatis serovar D was cloned into pQE30 vector following PCR amplification from genomic DNA. E. coli M15 transformants were induced to express the fusion protein by IPTG and the product was identified by SDS-PAGE and Western blot. Results: Confirmed by enzyme cleavage analysis and DNA sequencing, a correct recombinant plasmid pQE30/omp2 was constructed. The fusion protein from the transformants was approximately 60 kDa in size in SDS-PAGE analysis, which could specially react with anti-6 X His mouse monoclonal IgG antibodies. Conclusion: We successfully expressed Omp2 in E. coli M15, providing an efficient and simple system for assaying the immunological properties of Omp2.展开更多
文摘In order to compare the homology and antigen of Haemophilus parasuis (HPS)4 outer membrane protein P2(omp2), we design a test with specific primers, using PCR amplification of isolates of Haemophilus parasuis from Sichuan Province(HP Sch2010), ompP2 gene will be cloned into the pGM-T vector, and transformed into E. coli DH5α. Identified by PCR and sequencing and analysis, the sequencing results showed that the published 4 HPS SW124 strains omp2 gene (1077bp), compared with the amplified 1086bp purpose fragment(containing omp2 genes), is relatively stable, with the nucleotide homology level 97% and amino acid homology level of 92.5%. The variable regions are mainly concentrated in the three base sequences: 40-65,110-156,180-202.
基金This work was supported in part by grants from the Department of Science and Technology of Hunan Province (No. 01SSY2008-6) the Department of Health of Hunan Province (No. B2003-078).
文摘Objective: To construct a recombinant plasmid containing the outer membrane protein 2 (Omp2) gene of Chlamydia trachomatis and express Omp2 in E.coli. Methods: The omp2 gene of C. trachomatis serovar D was cloned into pQE30 vector following PCR amplification from genomic DNA. E. coli M15 transformants were induced to express the fusion protein by IPTG and the product was identified by SDS-PAGE and Western blot. Results: Confirmed by enzyme cleavage analysis and DNA sequencing, a correct recombinant plasmid pQE30/omp2 was constructed. The fusion protein from the transformants was approximately 60 kDa in size in SDS-PAGE analysis, which could specially react with anti-6 X His mouse monoclonal IgG antibodies. Conclusion: We successfully expressed Omp2 in E. coli M15, providing an efficient and simple system for assaying the immunological properties of Omp2.
