人朊蛋白基因真核表達(dá)載體的構(gòu)建及鑒定
發(fā)布時(shí)間：2018-12-30 19:53
【摘要】：目的構(gòu)建人朊蛋白基因(PRNP)真核表達(dá)重組質(zhì)粒。方法從阿爾茨海默病(AD)患者外周血白細(xì)胞中提取總RNA,利用RT-PCR的方法擴(kuò)增編碼人PRNP的基因片段,應(yīng)用基因重組技術(shù)將人PRNP基因片段克隆到真核表達(dá)載體pEGFP-N2中,經(jīng)XhoⅠ及EcoRⅠ雙酶切、單酶切及基因序列測定證實(shí)所構(gòu)建的載體。結(jié)果 RT-PCR方法獲得人PRNP的基因片段,限制性內(nèi)切酶酶切分析、PCR法及序列測定證實(shí)為正確重組質(zhì)粒,并且該重組載體能夠在SH-SY5Y細(xì)胞中表達(dá)。結(jié)論構(gòu)建的真核表達(dá)重組質(zhì)粒pEGFP-N2-PRNP通過鑒定,結(jié)構(gòu)正確,為后續(xù)研究PRNP的生物學(xué)功能奠定了基礎(chǔ)。
[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，
本文編號：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，
本文編號：2396059
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