人朊蛋白基因真核表达载体的构建及鉴定
发布时间：2018-12-30 19:53
【摘要】：目的构建人朊蛋白基因(PRNP)真核表达重组质粒。方法从阿尔茨海默病(AD)患者外周血白细胞中提取总RNA,利用RT-PCR的方法扩增编码人PRNP的基因片段,应用基因重组技术将人PRNP基因片段克隆到真核表达载体pEGFP-N2中,经XhoⅠ及EcoRⅠ双酶切、单酶切及基因序列测定证实所构建的载体。结果 RT-PCR方法获得人PRNP的基因片段,限制性内切酶酶切分析、PCR法及序列测定证实为正确重组质粒,并且该重组载体能够在SH-SY5Y细胞中表达。结论构建的真核表达重组质粒pEGFP-N2-PRNP通过鉴定,结构正确,为后续研究PRNP的生物学功能奠定了基础。
[Abstract]:Objective to construct the eukaryotic expression plasmid of human prion protein gene (PRNP). Methods Total RNA, was extracted from peripheral blood leukocytes of patients with Alzheimer's disease (AD). The gene fragment encoding human PRNP was amplified by RT-PCR and cloned into eukaryotic expression vector pEGFP-N2 by gene recombination technique. The vector was confirmed by Xho 鈪，
本文编号：2396059
[Abstract]:Objective to construct the eukaryotic expression plasmid of human prion protein gene (PRNP). Methods Total RNA, was extracted from peripheral blood leukocytes of patients with Alzheimer's disease (AD). The gene fragment encoding human PRNP was amplified by RT-PCR and cloned into eukaryotic expression vector pEGFP-N2 by gene recombination technique. The vector was confirmed by Xho 鈪，
本文编号：2396059
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