Expression of recombinant Rhipicephalus (Boophilus) microplus, R-annulatus and R-decoloratus Bm86 orthologs as secreted proteins in Pichia pastoris

被引：40

作者：

Canales, Mario ^{[1
]}

de la Lastra, Jose M. Perez ^{[1
]}

Naranjo, Victoria ^{[1
]}

Nijhof, Ard M. ^{[2
]}

Hope, Michelle ^{[3
]}

Jongejan, Frans ^{[2
,4
]}

de la Fuente, Jose ^{[1
,5
]}

机构：

[1] Inst Invest Recursos Cineget IREC CSIC UCLM JCCM, Ciudad Real 13071, Spain

[2] Univ Utrecht, Fac Vet Med, Dept Infect Dis & Immunol, UCTD, NL-3584 CL Utrecht, Netherlands

[3] CSIRO Livestock Ind, St Lucia, Qld 4067, Australia

[4] Univ Pretoria, Fac Vet Sci, Dept Vet Trop Dis, ZA-0110 Onderstepoort, South Africa

[5] Oklahoma State Univ, Dept Vet Pathobiol, Ctr Vet Hlth Sci, Stillwater, OK 74078 USA

来源：

BMC BIOTECHNOLOGY | 2008年 / 8卷 / 1期

基金：

英国惠康基金;

关键词：

D O I：

10.1186/1472-6750-8-14

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 [微生物学]; 0836 [生物工程]; 090102 [作物遗传育种]; 100705 [微生物与生化药学];

摘要：

Background: Rhipicephalus (Boophilus) spp. ticks economically impact on cattle production in Africa and other tropical and subtropical regions of the world. Tick vaccines constitute a cost-effective and environmentally friendly alternative to tick control. The R. microplus Bm86 protective antigen has been produced by recombinant DNA technology and shown to protect cattle against tick infestations. Results: In this study, the genes for Bm86 (R. microplus), Ba86 (R. annulatus) and Bd86 (R. decoloratus) were cloned and characterized from African or Asian tick strains and the recombinant proteins were secreted and purified from P. pastoris. The secretion of recombinant Bm86 ortholog proteins in P. pastoris allowed for a simple purification process rendering a final product with high recovery (35-42%) and purity (80-85%) and likely to result in a more reproducible conformation closely resembling the native protein. Rabbit immunization experiments with recombinant proteins showed immune cross-reactivity between Bm86 ortholog proteins. Conclusion: These experiments support the development and testing of vaccines containing recombinant Bm86, Ba86 and Bd86 secreted in P. pastoris for the control of tick infestations in Africa.

引用

页数：12

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