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OALib Journal期刊
ISSN: 2333-9721
费用：99美元
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干扰素诱导的跨膜蛋白-1基因的扩增、克隆及蛋白表达
, PP. 1221-1224
Keywords: 干扰素,跨膜蛋白-1基因,大肠肿瘤,pcr,克隆,蛋白表达
Abstract:
目的研究干扰素诱导的跨膜蛋白-1(interferon-inducibletransmembraneprotein-1,ifitmp-1)基因的扩增、克隆及蛋白表达。方法以含ifitmp-1基因的cdna为模板,用pfu酶做pcr扩增,用ecorⅰ和hindⅲ双酶切,将目的基因克隆到pucm-t质粒测序。进一步克隆到pet-trx蛋白表达载体质粒,优化蛋白表达条件。结果含有ifitmp-1基因的pcr产物约1000bp。重组pucm-t质粒经ecorⅰ、hindⅲ双酶切,有相应大小的cdna片段插入,测序结果显示其序列正确,证实目的基因的扩增、克隆成功,并成功克隆进pet-trx蛋白表达载体,经iptg诱导,在bl21(de3)plyss中有融合蛋白表达。结论成功扩增克隆ifitmp-1基因,并成功表达该基因编码的蛋白,为进一步研究ifitmp-1基因在大肠癌中的作用奠定了基础。
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