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。
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.
出处
《解剖学研究》
CAS
2007年第4期250-252,263,共4页
Anatomy Research
基金
广东省医学科研基金(NoB2006075)
