全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...

呼和浩特市区牛种布鲁菌bp26和omp10的克隆

DOI: 10.7668/hbnxb.2011.02.016, PP. 70-75

Keywords: 布鲁菌,bp2,omp10,同源,分析

Full-Text   Cite this paper   Add to My Lib

Abstract:

为明确试验布鲁菌和参考布鲁菌菌株与基因库中布鲁菌外膜蛋白BP2和OMP10基因间的同源性.利用布鲁菌外膜蛋白bp2和omp10基因,针对试验布鲁菌和参考布鲁菌菌株进行克隆和序列分析.根据GenBank发表的布鲁菌bp2基因和omp10基因,分别设计合成一对特异性引物,以提取的试验布鲁菌和2株参考布鲁菌的总DNA为模板,通过聚合酶链式反应(PCR)技术扩增得到bp2和omp10基因,回收纯化后将这两个基因分别连接到pMD18-T载体上,热激发转化受体菌大肠杆菌DH5a,质粒提取、PCR鉴定、测序.结果表明,bp2全长为995bp,包含一个由753bp组成的完整开放阅读框,试验菌株的同源性为100%,与参考菌株M5、S2的同源性分别为100%和99.9%,与参考序列S19、870的同源性分别为99.9%和100%.omp10全长531bp,包含一个由39bp组成的完整开放阅读框,试验菌株的同源性为100%,与参考菌株M5、S2的同源性均分别为100%,与参考序列544的同源性分别为99.7%.证实bp2基因和omp10基因在各种型间的同源性都在99%以上,表明这两个基因在布鲁菌属是高度保守的.

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.

Full-Text

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133