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周灵贵, 戴佳琳, 黄江, 廖兴江, 胡旭初, 余新炳, 申萍香, 郎书源. 亚洲带绦虫包虫诊断抗原P-29基因克隆表达[J]. 中国公共卫生, 2009, 25(9): 1091-1093. DOI: 10.11847/zgggws2009-25-09-38
引用本文: 周灵贵, 戴佳琳, 黄江, 廖兴江, 胡旭初, 余新炳, 申萍香, 郎书源. 亚洲带绦虫包虫诊断抗原P-29基因克隆表达[J]. 中国公共卫生, 2009, 25(9): 1091-1093. DOI: 10.11847/zgggws2009-25-09-38
ZHOU Ling-gui, DAI Jia-lin, HUANG Jiang, . Cloning,prokaryotic expression and immunoreacticity of hydatid disease diagnostic antigen P-29 of Taenia saginata asiatica[J]. Chinese Journal of Public Health, 2009, 25(9): 1091-1093. DOI: 10.11847/zgggws2009-25-09-38
Citation: ZHOU Ling-gui, DAI Jia-lin, HUANG Jiang, . Cloning,prokaryotic expression and immunoreacticity of hydatid disease diagnostic antigen P-29 of Taenia saginata asiatica[J]. Chinese Journal of Public Health, 2009, 25(9): 1091-1093. DOI: 10.11847/zgggws2009-25-09-38

亚洲带绦虫包虫诊断抗原P-29基因克隆表达

Cloning,prokaryotic expression and immunoreacticity of hydatid disease diagnostic antigen P-29 of Taenia saginata asiatica

  • 摘要: 目的 识别亚洲带绦虫包虫诊断抗原P-29(Hydatid disease diagnostic antigen P-29)的已知序列并行克隆和蛋白表达及免疫学研究.方法 利用在线生物信息学工具序列分析后以亚洲带绦虫成虫包虫诊断抗原P-29基因克隆到原核表达质粒pET-28a(+)中,在大肠埃希菌BL21/DE3中诱导表达,重组后的蛋白用His-镍蛋白纯化柱纯化,纯化的重组蛋白用蛋白印迹进行免疫学分析.结果 重组体构建成功并以包涵体形式存在,破包涵体纯化蛋白并得到高纯度的蛋白,且该重组蛋白可被亚洲带绦虫及牛带绦虫病人血清识别,表明其具有免疫反应性.结论 亚洲带绦虫成虫包虫诊断抗原P-29可在原核表达系统中获得具有免疫活性的高效表达.

     

    Abstract: Objective To identify the structure and function of hyda tid disease diagnosticantigen P229 of Taenia saginata asitica,and to construct prokaryotic recombinant plasmids of the gene,to express and purify the recom binant prote in and conduct a preliminary immunoreacticity study.Methods By on line geneanalys is at bio infomatics websites,the gene from the Taenia saginata as iatica full-length cDNA plasmid libratory was identified.The coding region sequence and the characteristics of the deduced prote in were analyzed.The recom binant prote in was detected by SDS-PAGE after being induced with IPTG in Ecoli BL 21/DE3 and purified with NI-IDA affinity chrom atography.Its immunoreactivity was determined by Western blotting.Results The recom binant plasmid was successfully constructed.The recombinant prote in could react with Ta enia asiatica and Taenia rhynchus saginatus infected patient's serum.Conclusion The hydatid disease diagnostic an tigen P 229 of Taenia saginata asitica was cloned and expressed,and the purified protein was confirmed with immunogenicity.

     

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