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绵羊FecB基因的原核表达及其抗体制备

DOI: 10.7668/hbnxb.2015.03.006, PP. 31-36

Keywords: FecB基因,原核表达,蛋白纯化,多克隆抗体制备

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Abstract:

为了进一步研究FecB基因,利用PCR-RFLP技术筛选出FecB基因,PCR扩增FecB基因编码区序列1509bp,利用限制性内切酶BamHⅠ和EcoRⅠ定向插入原核表达载体pET30a(+),构建原核表达载体pET30a(+)-FecB,诱导表达纯化重组蛋白,免疫小鼠制备多克隆抗体,WesternBlot检测重组蛋白的免疫活性,ELISA检测抗体效价。结果表明,成功的构建了绵羊FecB基因的原核表达载体,并在大肠杆菌中表达目的蛋白,经过4次免疫后,获得多克隆抗体,ELISA方法检测小鼠抗体免疫效价达到1:32000,纯化的蛋白具有免疫活性。为研究绵羊FecB基因蛋白的功能提供参考。

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