The Cloning of bp26 and omp1O Genes of Brucella Abortus in Huhhot

doi:10.7668/hbnxb.2011.02.016

Abstract

Abstract: To clear genes homology on gene of the Brucella outer membrane protein BP26 and OMPl0 genes a- mong Brucella of testing or reference and Genbank strains,clone and Comparison for their DNA sequences.We de. signed and synthesized a specific primers with Brucella 6p26 and omplO in Genbank.The genomic DNA of bp26 and omplO of testing or reference Brucella strains was extracted from each sample PCR amplification was then carried out and thus obtaining 6p26 and omplO.The PCR produce was inserted into pMDl8-T vector and sequenced.The plasmids were transformed into E.coli DH5a.Extracted E.coli plasmid DNA,PCR detected and sequenced.Sequencing showed that bp26 was 995 bp in length,including a753 bp open reading of the complete framework.Sequence comparison revealed that homology of bp26 among the testing strains was 100%,homology of bp26 of the testing and the reference M5 or S2 strains were 100%or 99.9%,homology of 6p26 of the testing strains and the reference S19 or 870 se. quences Was 99.9%or 100%.omplO Was 531 bp containing a396 bp open reading of the complete framework.homol- ogy of omplO among the testing strains was 100%homology of omplO of the testing and the reference M5 or S2 strains were 100%.homology of omplO of the testing and the reference 544 sequence were 997%There arebp26 and omplO gene homology between the various types in more than 99%.They is highly remain stable.

Key words: Brucella, bp26, omp10, Hanology, Analysis

CLC Number:

WANG Jun, WANG Rui, YANG Lian-ru, GUO Zhi-liang, YU Cui-ling, SHEN Zhi-yi. The Cloning of bp26 and omp1O Genes of Brucella Abortus in Huhhot[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2011, 26(2): 70-75. doi: 10.7668/hbnxb.2011.02.016.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2011.02.016

http://www.hbnxb.net/EN/Y2011/V26/I2/70

References

[1] Rossetti O L,Arese A I,Boschiroli M L,et al.Cloning of Brucella abortus gene and characterization of expressed 26-kilodalton periplasmic protein:potential use for diagnosis[J].J Clin Microbiol,1996,34(1):165-169.

[2] Salhi I,Bolgegrain R A,Machold J,et al.Characterization of new members of the group 3 outer membrane protein family of Brucella spp[J].Infect Immun,2003,71 (8):4326-4332.

[3] Seco-mediavilla P,Verger J M,Grayon M,et al.Epitope mapping of the Brucella melitensis bp26 immunogenic protein:usefulness for diagnosis of sheep brucellosis[J].Clin Diagn Lab Immunol,2003,10 (4):647-651.

[4] Yang X,Walters N,Robison A,et al.Nasal immunization with recombinant Brucella melitensis bp26 and trigger factor with cholera toxin reduces B.melitensis colonization[J].Vaccine,2007,25 (12):2261-2268.

[5] Cloeckanrt A,Jacques I,Grillo M J,et al.Development and evaluation as vaccines in mice of Brucella melitensis Rev.1 single and double deletion mutants of the bp26and omp31 genes coding for antigens of diagnostic significance in ovine brucellosis[J].Vaccine,2004,22 (21 -22):2827-2835.

[6] Debbarh H S,Zygmunt M S,Dubray (G,et al.Competitive enzyme-linked immunosorbent assay using monoclonal antibodies to the Brucella melitensis bp26 protein to evaluate antibody responses in infected and B.melitensis Rev.l vaccinated sheep[J].Vet Microbiol,1996,53 (3-4):325-337.

[7] Boschiroli M L,Cravero S L,Arese A I,et al.Protection against infection in mice vaccinated with a Brucella abortus mutant.Infect Immun,1997,65 (2):798-800.

[8] Tibor A,Saman E,de Wergifosse P,et al.Molecular characterization,occurrence,and immunogenicity in infected sheep and cattle of two minor outer membrane proteins of Brucella abortus[J].Infect lmmun,1996,64 (1):100 -107.

[9] Tibor A,Wansard V,Bielartz V,et al.Effect of omp10 or omp19 deletion on Brucella abortus outer membrane properties and virulence in mice[J].Infect Immun,2002,70(10):5540-5546.

[10] Cloeckaert A,Vizcaino N,Paquet J Y,et al.Major outer membrane proteins of Brucella spp.:past,present and future[J].Vet Microbiol,2002,90 (1-4):229-247.

[11] Cloeckaert A,Tibor A,Zygmunt M S.Brucella outer membrane lipoproteins share antigenic determinants with bacteria of the family Rhizobiaceae[J].Clin Diagn Lab Immunol,1999,6(4):627-629.

[12] Tibor A,Decelle B,Letesson J J.Outer membrane proteins Omp10,Omp16,and Omp19 of Brucella spp.are lipoproteins[J].Infect Immun,1999,67 (9):4960 -4962.

[13] Vizcaino N,Kittelberger R,Cloeckaert A,et al.Minor nucleotide substitutions in the omp31 gene of Brucella ovis result in antigenic differences in the major outer membrane protein that it encodes compared to those of the other Brucella species[J].Infect Immun,2001,69(11):7020-7028.

[14] 财音青格乐.基因工程及其产业[J].内蒙古农业科技,1999(S1):26-27,29.

[15] 高银.植物抗逆机制与基因工程研究进展[J].内蒙古农业科技,2007(5):75-78.

Please choose a citation manager

Content to export