重组质粒pEGFP-Brn-4的构建及其在骨髓间质干细胞中的表达  

Construction of recombinant plasmid pEGFP-Brn-4 and its expression in mesenchymal stem cells

作　　者：宣爱国[1] 龙大宏[1] 陈艳[2] 何峻峰[1] 

机构地区：[1]广州医学院人体解剖学教研室,广州510182 [2]广州医学院荔湾医院内科,广州510170

出　　处：《神经解剖学杂志》2008年第1期81-84,共4页Chinese Journal of Neuroanatomy

基　　金：广东省医学科研基金(NoB2006075)资助项目

摘　　要：本研究构建了真核表达重组质粒pEGFP-Brn-4,并观察了其在大鼠骨髓间质干细胞(MSC)中的表达情况。提取新生鼠脑组织总RNA,利用RT-PCR的方法扩增Brn-4的基因片段,用基因重组技术将Brn-4基因片段克隆到真核表达载体pEGFP-C2中,测序及PCR进行鉴定;将重组质粒应用脂质体转染的方法转入MSC中,荧光显微镜观察绿色荧光蛋白(GFP)的表达。经酶切及DNA测序结果证实pEGFP-Brn-4表达质粒的DNA序列完全正确,将此质粒转染MSC后Brn-4基因获得表达。研究结果提示成功构建真核表达重组质粒pEGFP-Brn-4,且Brn-4基因可在MSC中表达,为后续探讨Brn-4修饰的MSC治疗老年痴呆的可行性奠定了基础。The present study constructed an eukaryotic expression recombinant plasmid named pEGFP-Bru-4 and observed its expression in mesenchymal stem cells（MSC）. Total RNA was extracted from neogenetic rat brain and then reverse transcfiption-polymerase chain reaction （RT-PCR） tecnique was applied to amplify the Bru-4 gene. The product was inserted to eukaryotic expression vector pEGFP-C2 by using gene recombinant technique. We identified the new eukaryotic expression recombinant plasmid by PCR and DNA sequence analysis. Then pEGFP-Brn-4 was transfected into MSC by means of lipofectamine media methods. The results showed that the sequence of pEGFP- Brn-4 was identical with the genBank and the EGFP gene expression of Brn-4 can be observed in MSC after transfection. The results indicated that the recombinant plasmid pEGFP-Brn-4 was successfully constructed and Brn-4 gene can be observed in MSC. It will provide the bases for future study on therapy of Alzheimer disease.

关 键 词：Brn-4基因 重组质粒 骨髓间质干细胞 绿色荧光蛋白 

分 类 号：R346[医药卫生—基础医学]