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戴佳琳, 黄江, 廖兴江, 李波, 王宇, 江楠. 亚洲带绦虫EIFs3克隆与免疫反应性分析[J]. 中国公共卫生, 2010, 26(9): 1146-1148. DOI: 10.11847/zgggws2010-26-09-31
引用本文: 戴佳琳, 黄江, 廖兴江, 李波, 王宇, 江楠. 亚洲带绦虫EIFs3克隆与免疫反应性分析[J]. 中国公共卫生, 2010, 26(9): 1146-1148. DOI: 10.11847/zgggws2010-26-09-31
DAI Jia-lin, HUANG Jiang, LIAO Xing-jiang, . Cloning and immunoreactivity of full-length cDNA sequence of Taenia saginata asiatica eukaryotic translation initiation factor 3 subunit[J]. Chinese Journal of Public Health, 2010, 26(9): 1146-1148. DOI: 10.11847/zgggws2010-26-09-31
Citation: DAI Jia-lin, HUANG Jiang, LIAO Xing-jiang, . Cloning and immunoreactivity of full-length cDNA sequence of Taenia saginata asiatica eukaryotic translation initiation factor 3 subunit[J]. Chinese Journal of Public Health, 2010, 26(9): 1146-1148. DOI: 10.11847/zgggws2010-26-09-31

亚洲带绦虫EIFs3克隆与免疫反应性分析

Cloning and immunoreactivity of full-length cDNA sequence of Taenia saginata asiatica eukaryotic translation initiation factor 3 subunit

  • 摘要: 目的 对亚洲带绦虫真核翻译起始因子3(eukaryotic translation initiation factor3 subunit,EIFs 3)进行克隆及免疫学初步研究.方法 利用在线生物信息学工具从亚洲带绦虫成虫cDNA文库中筛选出含有EIFs3基因,并将该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,表达产物通过十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定,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(Western blotting)进行免疫学分析.结果 PCR,双酶切及DNA测序均显示重组体构建成功,用亲和层析法得到高纯度的蛋白,且该重组蛋白可被亚洲带绦虫及牛带绦虫及猪带病人血清识别,表明其具有免疫反应性.结论 亚洲带绦虫成虫EIFs 33基因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 clone and express the gene named as eukaryotic translation initiation factor 3 subunit(EIFs3)of Taenia saginata a sitica and to analyze the mimunogenicity of its recombinant protein.Methods By online analysis of the gene at bioinfom aticswebsites,the gene from the Taenia saginata a siatica full length cDNA plasmid libratory was identi fied and its coding region sequence was analyzed.Then the coding region of the gene was amplified with PCR.The recombinant prote in was detected by sodium dodecyl sulfate polyacry lamide gelelec trophoresis(SDS PAGE)after being induced with isopro pyl thiogalacto side(IPTG)in Ecoli BL 21/DE3 and purified with NI IDA affinity chroma to graphy.In addition,the immuno reactivity of the purified recombinant prote ins was analyzed with We stern blotting.Results As demonstrated by PCR,double enzyme digestion and DNA sequencing of the recom binant plasmid was successively constructed.SDS PAGE results showed that the gene was expressed in E coli BL 21/DE3 and the recom binant prote in could react with Taenia a siatica andtae niarhynchus saginatus infected patientse rum,which indicated its mimuno reacticity.Conclusion The EIFs 3 of Taeniasaginata a sitica was cloned and expressed,and the purified protein has mimunogeni city.

     

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