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| | Objective: To construct the recombinant adenovirus 5 shuttle vector with NR2B gene.Methods:NR2B gene was obtained by digesting the plasmids pcDNA3.1-NR2B with(EcoRⅠ)and(BamHⅠ) and was cloned into the adenovirus shuttle vector pDC515.Then the vector was transferred into 293 cell and the expression of NR2B in 293 cell was identified by RT-PCR and Western blot.Results:The restriction endonuclease digestion and DNA sequencing confirmed that NR2B recombined adenovirus shuttle vector was constructed successfully.... | | 目的:构建携带N-甲基-D天冬氨酸(NMDA)受体2B亚基(NR2B)基因的腺病毒5型(Ad5)穿梭载体,为NR2B重组腺病毒镇痛疫苗的包装做准备。方法:以限制性内切酶EcoRⅠ和BamHⅠ双酶切质粒pcDNA3.1-NR2B获得目的基因NR2B;将NR2B定向克隆入腺病毒穿梭载体pDC515,并行酶切及DNA测序鉴定。在脂质体介导下将pDC515-NR2B转染293细胞,并以RT-PCR及W estern b lot分别从转录水平和翻译水平检测目的基因NR2B的表达。结果:经酶切和DNA测序证实目的基因NR2B被成功克隆入载体pDC515-NR2B中,RT-PCR及W estern b lot检测NR2B可在293细胞表达。结论:成功构建NR2B重组Ad5穿梭载体pDC515-NR2B,该载体可用于NR2B重组Ad5载体镇痛疫苗的包装。 | | 文摘来源 | |   |
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