摘要
目的以pEGFP-N1质粒载体为介导,构建针对髓鞘碱性蛋白(MBP)不同isoform cDNA的表达载体。方法将三段不同MBPcDNA序列克隆入pEGFP-N1质粒,转化DH5α感受态细胞,酶切鉴定及测序。结果测序结果显示插入的三段目的片段的序列与理论序列一致。结论笔者成功构建了pEGFP-N1-MBP21.5,pEGFP-N1-MBP18.5和pEGFP-N1-MBP17.3三个重组真核表达载体,为进一步研究不同MBP cDNA在真核细胞中的表达情况和功能差异奠定基础。
Objective To construct different expression vectors of isoform cDNA aiming at MBP,taking pEGFP-N1 plasmid as mediation.Methods Three different MBP cDNA sequences were cloned into pEGFP-N1 plasmid and transformed into DH5α competent cells,which were identified and screened by restriction enzyme analysis.Results The sequencing results showed that the sequences of three inserted purposed segments were the same as theory sequences.Conclusion We construct three recombinant eukaryotic expression vectors successfully：pEGFP-N1-MBP21.5,pEGFP-N1-MBP18.5 and pEGFP-N1-MBP17.3,which lay a foundation for the further study of the expression and functional differences of different MBP cDNA in eukaryotic cells.
出处
《中国当代医药》
2012年第36期5-6,9,共3页
China Modern Medicine
基金
四川省教育厅重点项目(07ZA029)
