重组质粒pEGFP-Brn-4的构建与鉴定  

Construction and identification of recombinant plasmid pEGFP-Brn-4

作　　者：陈艳[1] 方继荣[1] 宣爱国[2] 

机构地区：[1]广州医学院荔湾医院,广州510170 [2]广州医学院解剖教研室

出　　处：《临床医学》2011年第1期99-100,共2页Clinical Medicine

基　　金：广州市医药卫生科技项目(2009-YB-174)

摘　　要：目的构建神经同源盒基因Brn-4真核表达重组质粒。方法从新生SD大鼠脑组织提取总RNA,利用逆转录聚合酶链反应(RT-PCR)的方法扩增编码鼠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,经BamHI、EcoRⅠ双酶切和单酶切证实所构建的载体。结果 RT-PCR方法获得鼠Brn-4的基因片段,限制性内切酶酶切分析和PCR法鉴定表明为正确重组子。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,为后续研究在神经干细胞中表达及其体内研究奠定了基础。Objective To construct an eukaryotic expression recombinant plasmid named pEGFP-Brn-4.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 cDNA was inserted into retroviral vector pEGFP-C2.The recombinant plasmid was identified by a pair of specified primers containing the restriction sites of BamH I and EcoRⅠ.Results The Brn-4 gene could be obtained by RT-PCR,the recombinant plasmid was identified by restriction endonuclease analysis and PCR.Conclusion The recombinant plasmid pEGFP-Brn-4 is constructed successfully.Which offers a help for the further research on the expression in neural stem cells and the study in vivo.

关 键 词：Brn-4基因 真核表达载体 基因重组 

分 类 号：R346[医药卫生—基础医学]