人单纯疱疹病毒BDNF转基因重组体的构建表达及活性检测

Constrution of Human Single Herpes Virus Carried BDNF Gene Vector and Its Bioactivity Evaluation

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。

Objective To construct the recombinant vector of human single herpes virus（HSV） carried brain derived neurotrophic factor（BDNF） gene.Methods BDNF gene was acquired from rat brain tissue by RT-PCR,then was cloned into plasmid,and enveloped by HSV.The recombinant was used to transfer cultured cortical neurons.The number and neurite length of neurons were quantified.The BDNF level and subcellular localization were detected by ELISA and immunohistochemistry.Results HSV carried BDNF gene recombinant has been successfully constructed.The recombinant showed the bioactivity on the growth of cortical neurons.BDNF level was increased significantly in BDNF transferred group.Conclusion HSV carried BDNF gene recombinant,with the bioactivity,has been successfully constructed.This could provide the vector for the treatment of BDNF under disease condition base on transferring gene technique.