Bdnf基因过表达慢病毒载体的构建及表达*

doi:10.12047/j.cjap.6280.2022.074

中国应用生理学杂志 ›› 2022, Vol. 38 ›› Issue (5): 397-400.doi: 10.12047/j.cjap.6280.2022.074

Bdnf基因过表达慢病毒载体的构建及表达^*

钟锦, 史晋朝, 张倩, 孟金凤, 张倩倩, 李建国^△

山西医科大学生理学系, 细胞生理学教育部重点实验室, 太原 030001

收稿日期:2022-02-25 修回日期:2022-09-28 出版日期:2022-09-28 发布日期:2023-04-23
通讯作者: ^△Tel: 13935128325; E-mail: lijg@sxmu.edu.cn
基金资助:
^*山西省基础研究计划面上项目(20210302123305)

Construction and expression of BDNF gene overexpression lentiviral vector

ZHONG Jin, SHI Jin-chao, ZHANG Qian, MENG Jin-feng, ZHANG Qian-qian, LI Jian-guo^△

Received:2022-02-25 Revised:2022-09-28 Online:2022-09-28 Published:2023-04-23

摘要/Abstract

摘要： 目的: 探讨脑源性神经营养因子(BDNF)基因过表达慢病毒载体的构建和包装,检测其在大鼠海马原代神经元中的表达。方法: 利用聚合酶链式反应(PCR)技术扩增Bdnf基因的exons4和CDS序列,将其克隆到pcDNA3.1-mCherry载体上,构建pcDNA3.1-exons4-Bdnf-mCherry慢病毒表达载体。重组质粒经KpnI和EcoRI双酶切鉴定和测序验证。使用三质粒包装系统将重组质粒pcDNA3.1-exons4-Bdnf-mCherry和慢病毒辅助包装质粒pMDLg/pRRE、pRSV-Rev和pCMV-VSV-G共转染293T细胞,制备并浓缩慢病毒颗粒,感染大鼠原代海马神经元,荧光显微镜和实时荧光定量PCR检测目的基因在原代海马神经元中的表达。结果: 成功构建pcDNA3.1-exons4-Bdbf-mCherry慢病毒过表达载体,转染后的293T细胞表达红色荧光,并在培养大鼠海马原代神经元中表达增加(P＜0.05)。结论: 成功构建含有特定转录本的Bdnf基因过表达慢病毒载体,为进一步深入研究Bdnf的作用提供技术基础。

关键词: 脑源性神经营养因子, 慢病毒, 293T细胞, 原代海马神经元, 基因表达, 细胞培养

Key words: brain-derived neurotrophic factor, lentivirus, 293T cells, primary hippocampal neurons, gene expression, cell cultures

中图分类号:

Q189

钟锦, 史晋朝, 张倩, 孟金凤, 张倩倩, 李建国. Bdnf基因过表达慢病毒载体的构建及表达^*[J]. 中国应用生理学杂志, 2022, 38(5): 397-400.

ZHONG Jin, SHI Jin-chao, ZHANG Qian, MENG Jin-feng, ZHANG Qian-qian, LI Jian-guo. Construction and expression of BDNF gene overexpression lentiviral vector[J]. CJAP, 2022, 38(5): 397-400.

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Bdnf基因过表达慢病毒载体的构建及表达^*

Construction and expression of BDNF gene overexpression lentiviral vector

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