人单纯疱疹病毒BDNF转基因重组体的构建表达及活性检测
Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition(2012)
Abstract
目的 构建人单纯疱疹病毒(HSV)脑源性神经营养因子(BDNF)转基因重组体(HSV-BDNF),转染大鼠皮质神经元后并检测其表达及生物活性.方法 用RT-PCR方法扩增人BDNF基因,克隆并构建HSV-BDNF转基因重组体.构建成功后将其转染培养大鼠皮质神经元,观察其对皮质神经元存活的影响;用ELISA、免疫组化检测皮质神经元中BDNF的表达水平.结果 成功构建了HSV-BDNF转基因重组体,皮质神经元转染HSV-BDNF后BDNF表达水平明显上调(P<0.05),且明显促进皮质神经元生长(P<0.05).结论 成功构建了HSV-BDNF,能有效转染大鼠皮质神经元并促进BDNF高表达,且具有生物活性.
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Key words
Brain derived neurotrophic factor,Cortical neuron,Gene cloning,Human single herpes virus,Transfection
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