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首页> 外文期刊>Research in Veterinary Science >Cloning, expression, and characterization of the MSP1a and MSP1b recombinant proteins from PR1 Anaplasma marginale strain, Brazil
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Cloning, expression, and characterization of the MSP1a and MSP1b recombinant proteins from PR1 Anaplasma marginale strain, Brazil

机译：来自巴西PR1 Anaplasma marginale菌株的MSP1a和MSP1b重组蛋白的克隆，表达和表征

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摘要

The Anaplasma marginale is a bacterium that has obligate intraerythrocytic multiplication in cattle causing important economic loss. The A. marginale major surface protein 1 (MSP1) complex, heterodimer composed of MSP1a and MSP1b, has been identified as adhesins for bovine erythrocytes. The objectives of this study were to sequences the msp1beta gene and produce and characterize recombinant MSP1a and MSP1b from a Brazilian strain of A. marginale, PR1. The msp1alpha and msp1beta genes from the PR1 strain were cloned and expressed in E. coli BL21 Star using the vectors pET102 and pET101/D-TOPO. Antibodies were produced against the recombinant proteins and were shown to react with rMSP1a and rMSP1b demonstrating a molecular mass of 70kDa to 105kDa and 100kDa, respectively for these proteins. Bovine erythrocytes were agglutinated by BL21/rMSP1a and BL21/rMSP1b and, this agglutination was inhibited by the presence of the IgY anti-rMSP1a, confirming the adhesion function of these proteins. Additionally, using the IgY anti-rMSP1a and rMSP1b in a IFI, the presence of rMSP1a and rMSP1b was confirmed on the outer membrane of the recombinant E. coli BL21. Our results show that the msp1beta gene from the PR1 strain has both the conserved region and contain the defined polymorphism regions previously described for other strains of A. marginale. The results from this study confirm adhesive functions for rMSP1a and rMSP1b from PR1 strain in bovine erythrocytes invasion.
机译：Anaplasma marginale是一种细菌，在牛中具有专一性的红细胞内繁殖，造成重大的经济损失。由MSP1a和MSP1b组成的异质二聚体A.marginale主要表面蛋白1（MSP1）复合物已被鉴定为牛红细胞的粘附素。这项研究的目的是对msp1beta基因进行测序，并从巴西拟南芥PR1株中生产并鉴定重组MSP1a和MSP1b。使用载体pET102和pET101 / D-TOPO克隆PR1株的msp1alpha和msp1beta基因并在大肠杆菌BL21 Star中表达。产生了针对重组蛋白的抗体，并显示它们与rMSP1a和rMSP1b反应，表明这些蛋白的分子量分别为70kDa至105kDa和100kDa。牛红细胞被BL21 / rMSP1a和BL21 / rMSP1b凝集，这种凝集被IgY抗rMSP1a的存在抑制，证实了这些蛋白的粘附功能。此外，在IFI中使用IgY抗rMSP1a和rMSP1b，可在重组大肠杆菌BL21的外膜上确认rMSP1a和rMSP1b的存在。我们的结果表明，来自PR1菌株的msp1beta基因既具有保守区域，又包含先前针对其他边缘拟南芥菌株描述的定义的多态性区域。这项研究的结果证实了PR1株rMSP1a和rMSP1b在牛红细胞入侵中的黏附功能。

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