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首页> 外文期刊>Infection and immunity >Properties of pertussis toxin B oligomer assembled in vitro from recombinant polypeptides produced by Escherichia coli.
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Properties of pertussis toxin B oligomer assembled in vitro from recombinant polypeptides produced by Escherichia coli.

机译：百日咳毒素B寡聚体的性质是由大肠杆菌生产的重组多肽体外组装而成。

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摘要

The subunits that make up the pentameric B oligomer of pertussis toxin (S2, S3, S4, and S5) were individually synthesized as recombinant polypeptides in Escherichia coli, isolated as insoluble inclusion bodies, and assembled into a multimeric form in vitro by spontaneous association following treatment with a chaotropic agent, reduction, and reoxidation. The recombinant B multimer, purified by fetuin-Sepharose affinity chromatography, contained all four of the individual subunits and possessed the mitogenic and hemagglutinating activities characteristic of the native B oligomer. Immunization of mice with the recombinant B oligomer elicited antibodies that neutralized pertussis toxin in vitro and, moreover, provided protection in vivo against the leukocytosis-promoting activity of the toxin. These results demonstrate the potential for assembly of complex multimeric proteins from recombinant DNA-derived polypeptides and provide a novel means for production of an acellular pertussis vaccine component.
机译：组成百日咳毒素五聚体B寡聚体的亚基（S2，S3，S4和S5）在大肠杆菌中分别作为重组多肽合成，分离为不溶性包涵体，并在体外通过以下自发结合组装成多聚体形式用离液剂处理，还原和再氧化。通过胎球蛋白-琼脂糖亲和色谱纯化的重组B多聚体包含所有四个单独的亚基，并具有天然B寡聚体的促有丝分裂和血凝活性。用重组B寡聚体对小鼠进行免疫引发了抗体，该抗体在体外中和了百日咳毒素，并且在体内提供了针对毒素的促进白细胞增多的活性的保护作用。这些结果证明了从重组DNA衍生的多肽组装复杂的多聚体蛋白质的潜力，并提供了生产脱细胞百日咳疫苗组分的新颖手段。

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