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大豆科学  2009 

大豆主要过敏原GlymBd30K基因的克隆及其原核表达载体的构建

DOI: 10.11861/j.issn.1000-9841.2009.01.0011, PP. 11-15

Keywords: 大豆,过敏原,GlymBdK,克隆,原核表达载体

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Abstract:

利用RT-PCR克隆大豆主要过敏原GlymBd30K的全长基因,根据序列设计带有酶切位点的特异性引物,扩增大豆GlymBd30K的完整开放阅读框,与pET-28a载体连接,构建原核表达载体。结果表明:克隆了大豆主要过敏原GlymBd30K基因,且构建了其原核表达载体。该基因含有长度为1140bp的开放阅读框,编码379个氨基酸。该蛋白质的相对分子质量为42758,等电点为5.08。序列同源性分析发现其与数据库中已知的GlymBd30K基因同源性很高,因此认为其是大豆的过敏原基因,在GenBank数据库中的登录号为EU883600。克隆的大豆主要过敏原GlymBd30K基因及构建的原核表达载体,为大豆主要过敏原GlymBd30K的重组表达和免疫活性鉴定等奠定基础。

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