| PDZ1-腺病毒重组体的构建及其对KA诱导的海马神经元凋亡的拮抗作用 |
| |
| 引用本文： | 胡书群,纵艳艳,侯筱宇,张光毅.PDZ1-腺病毒重组体的构建及其对KA诱导的海马神经元凋亡的拮抗作用[J].免疫学杂志,2008,24(6):651-655. |
| |
| 作者姓名： | 胡书群 纵艳艳 侯筱宇 张光毅 |
| |
| 作者单位： | 徐州医学院生物化学与分子生物学研究中心江苏省脑病生物信息重点实验室; |
| |
| 基金项目： | 国家自然科学基金(30330190);;江苏省高校自然科学基础研究基金(07KJB310118);;徐州医学院科研基金(07KJ15)资助课题 |
| |
| 摘 要： | 目的构建PSD-95结构域PDZ1与腺病毒重组体并观察其对海人藻酸(KA)诱导的海马神经元凋亡的作用。方法采用RT-PCR法获得PDZ1的cDNA;与腺病毒穿梭载体pAdTrack-CMV用T4 DNA连接酶连接;连接产物与骨架载体pAdEasy-1共转染至原核包装细胞BJ5183;将原核重组体用脂质体转入真核包装细胞293H;纯化包装后的病毒颗粒并测定其滴度;腺病毒感染培养的海马神经元,加入KA,用DAPI染色后荧光显微镜观察细胞凋亡;用流式细胞仪定量分析细胞凋亡率。结果①经RT-PCR得到了PDZ1的cDNA;②电泳鉴定得到原核重组体;③得到包装完整的具感染能力的病毒颗粒;④纯化的病毒颗粒滴度为1.08×109ifu/mL⑤病毒表达的PDZ1使KA诱导的海马神经元凋亡数量减少(P<0.05)。结论获得了能表达PDZ1-GFP的病毒颗粒,并且PDZ1过表达能拮抗KA诱导的海马神经元凋亡。
|
| 关 键 词： | PDZ1 腺病毒载体 KA 海马神经元 凋亡 |
Construction of recombinant Ad-PDZ1-GFP and its antagonistic effects on kainic acid-induced apoptosis of hippocampl neurons |
| |
| HU Shu-qun,ZONG Yan-yan,HOU Xiao-yu,ZHANG Guang-yi.Construction of recombinant Ad-PDZ1-GFP and its antagonistic effects on kainic acid-induced apoptosis of hippocampl neurons[J].Immunological Journal,2008,24(6):651-655. |
| |
| Authors: | HU Shu-qun ZONG Yan-yan HOU Xiao-yu ZHANG Guang-yi |
| |
| Institution: | Research Center of Biochemistry and Molecular Biology;Xuzhou Medical College;Xuzhou 221002;China |
| |
| Abstract: | Objective To construct recombinant of Ad-PDZ1-GFP and detect its effect on apoptosis in neurons induced by kainic acid (KA). Methods Using the total RNA as template,the cDNA of PDZ1 was amplified by RT-PCR; the cDNA was subsequently cloned into the shuttle vector pAdTrack-CMV; the resultant plasmid is co-transformed into E. coli BJ5183 cells with an adenoviral backbone plasmid pAdEasy-1; the recombinant plasmid is transfected into adenovirus packaging cell lines HEK 293 cells with Lipofectamine; the particl... |
| |
| Keywords: | PDZ1 KA |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
