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溶藻弧菌转运蛋白TolB的免疫原性与免疫保护性

DOI: 10.13560/j.cnki.biotech.bull.1985.2015.07.021, PP. 143-149

Keywords: 溶藻弧菌,tolB基因,免疫原性,免疫保护

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Abstract:

旨在研究溶藻弧菌(Vibrioalginolyticus)HY9901转运蛋白TolB作为疫苗候选抗原的可能性,根据已发表的全基因组序列,设计特异性引物PCR扩增tolB的基因全长序列。序列分析显示,该基因(GenBank登录号JQ846501)全长1353bp,共编码450个氨基酸残基。BLAST分析发现,溶藻弧菌tolB基因与其他已知弧菌的tolB具有较高的同源性,序列保守,可作为共同抗原候选蛋白。用原核表达的tolB蛋白免疫SPF级小鼠,制备多克隆抗体,ELISA效价达1∶40000。免疫印迹表明鼠抗tolB血清能与诱导后的重组蛋白发生特异反应。为进一步研究tolB对石斑鱼(Epinephelusawoara)的免疫保护性,用TolB蛋白两次免疫石斑鱼,ELISA检测发现,免疫石斑鱼血清的抗体效价在第4周达到峰值(1∶2048)。攻毒实验结果表明TolB对石斑鱼的免疫保护率为76%。结果表明,溶藻弧菌转运蛋白TolB具有较好的免疫原性和免疫保护性,可作为弧菌亚单位疫苗的候选抗原。

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