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Position: Home > Articles > Prokaryotic Expression and Antibody Preparation of SdrD Surface Protein of Staphylococcus aureus Progress in Veterinary Medicine 2015 (3) 1-5

金黄色葡萄球菌表面蛋白SdrD的原核表达及抗体制备

作  者:
杜欣军;王学连;李萍;耿洁洁;张洪娟;王硕
单  位:
食品营养与安全教育部重点实验室;天津科技大学食品工程与生物技术学院
关键词:
金黄色葡萄球菌;表面蛋白;SdrD基因;重组表达;抗体制备
摘  要:
根据金黄色葡萄球菌表面蛋白SdrD基因序列设计特异性引物,以金黄色葡萄球菌标准菌株基因组为模板进行PCR扩增,获得了大小为2 043bp的基因片段。将基因片段连接至表达载体pET26b,转入大肠埃希菌BL21(DE3)中用IPTG诱导表达。SDS-PAGE结果表明,SdrD基因获得了较好的表达,重组蛋白的分子质量约为99ku。利用免疫亲和层析对目标蛋白进行纯化,获得了纯度较高的目标蛋白。用该蛋白免疫新西兰白兔制备了多克隆抗体,抗体效价达到1∶23 000,为金黄色葡萄球菌基因工程疫苗的开发与性能评价,以及该致病菌免疫检测技术的开发奠定了基础。
译  名:
Prokaryotic Expression and Antibody Preparation of SdrD Surface Protein of Staphylococcus aureus
作  者:
DU Xin-jun;WANG Xue-lian;LI Ping;GENG Jie-jie;ZHANG Hong-juan;WANG Shuo;Key Laboratory of Food Nutrition and Safety,Ministry of Education,Tianjin University of Science & Technology;
关键词:
Staphylococcus aureus;;surface protein;;SdrD gene;;recombinant expression;;antibody preparation
摘  要:
In this study,specific primers were designed and synthesized based on the SdrD gene of Staphylococcus aureus and used to amplify the gene fragment using the genomic DNA of S.aureus standard strain as template.A gene fragment of 2 043 bp in length was obtained.The ORF was ligated into pET26 bexpression vector and transformed into E.coli BL21(DE3)for recombinant expression under induction of IPTG.The results of SDS-PAGE indicated that the recombinant protein with molecular weight of 99 ku was successfully expressed.The target protein was purified using immunoaffinity chromatography and used to immunize New Zealand white rabbits to prepare polyclonal antibodies.ELISA demonstrated that the titers of antiserum reached 1∶23 000.This study lays a foundation for developing engineered vaccines and immune detection methods for S.aureus.

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