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美洲大蠊Per a7基因的克隆、表达及免疫学鉴定
  • 期刊名称:刘志刚,何毅华,吴海强,朱永峰,美洲大蠊Per a7基因的克隆、表达及免疫学鉴定,昆虫学报, 50(
  • 时间:0
  • 分类:Q966[生物学—昆虫学]
  • 作者机构:[1]深圳大学生命科学学院,广东深圳518060
  • 相关基金:国家自然科学基金项目(39860071,30571625);广东省科技计划重点项目(2003A3080502);深圳市科技计划项目(200326)
  • 相关项目:蟑螂重组低致敏原在哮喘特异性免疫治疗中的机理研究
中文摘要:

根据原肌球蛋白基因序列设计引物,以我国南方地区美洲大蠊Periplaneta americana RNA为模板,用RT-PCR方法扩增出852bp的全长编码片段,经序列分析发现该基因与NCBI公布的Pera7有3个核苷酸发生改变。将目的片段克隆到pET24a(+)表达载体中,在大肠杆菌BL21Star获得表达,融合蛋白的分子量约为33kD。利用蟑螂过敏性患者血清对表达产物进行Western blotting检测,出现明显的识别条带,说明表达产物具有IgE结合活性。

英文摘要:

An 852 bp cDNA fragment was amplified by RT-PCR from total mRNA of the American cockroach, Periplaneta americana (South China population) with a pair of primers that were designed according to the published tropomyosin gene. Sequence analysis indicated that three nucleotides changed in the gene compared with the reported Per a7 gene. The fragment was cloned into expression vector pET24a( + ) and subsequently expressed in E. coli BL21 Star. SDS-PAGE analysis showed that the molecular weight of the expressed product was 33 kD. Western blotting indicated that the recombinant protein could combine with sera IgE from human individuals allergic to cockroach allergens.

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