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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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人arpc2基因的克隆与表达
Keywords: arpc2,载体构建,基因表达,蛋白纯化
Abstract:
目的构建人his-arpc2融合蛋白表达载体,并在原核细胞中进行表达与纯化,以研究其功能及鉴定与其相互作用的蛋白。方法采用pcr方法从人肝cdna文库扩增arpc2基因编码区,使用常规酶切、连接方法将其重组至pet-14b载体中。对阳性克隆进行酶切、pcr和测序鉴定。转化bl21(de3)大肠杆菌株,用异丙基β-d硫代半乳糖(iptg)诱导融合蛋白表达,并利用ni-nta亲和层析纯化该融合蛋白。结果所构建的人arpc2的his融合蛋白表达载体是正确的,该载体可在大肠杆菌内高效表达,用ni-nta纯化获得了相对分子质量约36000的融合蛋白。结论成功构建了人his-arpc2融合蛋白表达载体,并在原核细胞中获得了高效表达和纯化,为进一步研究arpc2的生理功能提供了重要的实验材料。
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