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重组微小扇头蜱（牛蜱属）、环形扇头蜱和无色扇头蜱Bm86直系同源物作为分泌蛋白在毕赤酵母中的表达。

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

作者信息

Canales Mario, de la Lastra José M Pérez, Naranjo Victoria, Nijhof Ard M, Hope Michelle, Jongejan Frans, de la Fuente José

机构信息

Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), Ronda de Toledo s/n, 13071 Ciudad Real, Spain.

出版信息

BMC Biotechnol. 2008 Feb 14;8:14. doi: 10.1186/1472-6750-8-14.

DOI:10.1186/1472-6750-8-14
PMID:18275601
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2262073/
Abstract

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.

摘要

背景

璃眼蜱属（牛蜱属）蜱虫对非洲以及世界其他热带和亚热带地区的养牛业造成经济影响。蜱虫疫苗是一种经济高效且环保的蜱虫控制替代方法。微小牛蜱的Bm86保护性抗原已通过重组DNA技术生产出来，并显示出能保护牛免受蜱虫侵扰。

结果

在本研究中，从非洲或亚洲蜱虫菌株中克隆并鉴定了Bm86（微小牛蜱）、Ba86（环形牛蜱）和Bd86（变异革蜱）的基因，重组蛋白从巴斯德毕赤酵母中分泌并纯化出来。重组Bm86直系同源蛋白在巴斯德毕赤酵母中的分泌使得纯化过程简单，最终产品回收率高（35 - 42%）、纯度高（80 - 85%），并且可能产生更可重复的构象，与天然蛋白非常相似。用重组蛋白进行的兔子免疫实验表明Bm86直系同源蛋白之间存在免疫交叉反应。

结论

这些实验支持开发和测试含有在巴斯德毕赤酵母中分泌的重组Bm86、Ba86和Bd86的疫苗，用于控制非洲的蜱虫侵扰。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/f0eb1a2f3585/1472-6750-8-14-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/7022d40b3ddc/1472-6750-8-14-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/045c5fe20fb5/1472-6750-8-14-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/54a5408d6ed4/1472-6750-8-14-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/bfeecfacc9d9/1472-6750-8-14-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/f0eb1a2f3585/1472-6750-8-14-5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/7022d40b3ddc/1472-6750-8-14-1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/045c5fe20fb5/1472-6750-8-14-2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/54a5408d6ed4/1472-6750-8-14-3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/bfeecfacc9d9/1472-6750-8-14-4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d25f/2262073/f0eb1a2f3585/1472-6750-8-14-5.jpg

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