学术
重组质粒pEGFP—Brn-4的构建及其在神经干细胞中的表达
宣爱国[1] 龙大宏[1] 何峻峰[1] 陈艳[2]
[1]广州医学院人体解剖教研室,广东广州510182 [2]广州医学院荔湾医院,广东广州510182
摘 要:
目的构建神经发育转录因子Brn-4基因真核表达重组质粒，研究Brn-4在神经干细胞（NSC）中的表达情况。方法从新生鼠脑组织提取总RNA，利用RT—PCR的方法获得编码鼠Brn-4的基因片段，应用基因重组技术，将鼠Brn-4基因片段克隆到真核表达载体pEGFP—C2中．应用脂质体转染NSC．荧光显微镜观察该Brn-4基因在NSC中的表达。结果限制性内切酶酶切分析和PCR法鉴定表明为正确重组子，Brn-4基因在NSC中获得表达。结论新构建的真核表达重组质粒pEGFP—Brn-4通过鉴定，结构正确；Brn-4基因可在NSC中表达。可作为后续研究老年性痴呆动物模型转基因实验的基因来源。[著者文摘]
文章出处:
《解剖学研究》-2007年29卷4期,263 -250-252,263页
栏目信息:
分 类 号:
Construction and expression of recombinant plasmid pEGFP-Brn-4 in neural stem cells
XUAN Ai-guo, LONG Da-hong, HE Jun-feng, CHEN Yan（ Department of Anatomy, Guangzhou Medical College, Guangzhou 510182 China ）
Abstract:
Objective To construct an eukaryotic expression recombinant plasmid named pEGFP-Brn-4, and research its expression in neural stem cells （NSC）. Methods Total RNA was extracted from rat brain as the template and the Brn-4 gene was amplified by reverse transcription-polymerase chain reaction （RT-PCR）. By using gene recombination technique, rat Brn-4 eDNA was inserted into eukaryotic expression vector pEGFP-C2. And then, the NSC were transferred by means of lipofectamine media methods. After that, we observed the Brn-4 expressed by NSC. Results The recombinant plasmid was identified to be fight by restriction endonuclease analysis and PCR. The Brn-4 gene could be expressed successfully by NSC. Conclusion The recombinant plasmid pEGFP-Brn-4 is constructed successfully. The Brn-4 gene can be expressed successfully by NSC, which offers a help on gene therapy of Alzheimer disease.[著者文摘]
Key words:
Brn-4 gene; Eukaryotic expression vector; Neural stem cell; Green fluorescent protein
基金资助:
广东省医学科研基金（No.B2006075）
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