摘要
目的应用pcDNA3.1(-)质粒载体系统构建携带小鼠铁蛋白重链全基因质粒,并探讨铁蛋白基因作为一种新型内源性磁共振分子影像报告基因的可行性和优越性。方法从来源于小鼠肌肉的mRNA,应用一对含有限制性内切酶NotⅠ和BamHⅠ酶切位点的引物进行逆转录聚合酶链反应(RT-PCR),扩增出铁蛋白重链全基因(Fth)片断。通过限制性内切酶NotⅠ和BamHⅠ酶切和T4连接酶的作用,将铁蛋白重链全基因插入到载体pcDNA3.1(-)中,构建铁蛋白重链全基因质粒pcDNA3.1(-)-Fth。结果质粒pcDNA3.1(-)-Fth成功构建,符合酶切和测序鉴定结果。结论初步构建的小鼠铁蛋白重链质粒,为进一步探讨铁蛋白基因作为一种新型内源性磁共振分子影像报告基因在细胞示综和基因显像中的应用奠定了基础。
Objective To construct the plasmid of heavy chain of mice ferritin, and to research its possibility and superiority of being a new endogenous MRI reporter-ferritin. Methods The fragment of heavy chain of ferritin （Fth） was amplified from murine muscle mRNA using RT-PCR with a set of primers containing digestive sequences of restriction enzyme Not 1 and BamH Ⅰ. The Fth fragment was inserted into pcDNA3.1 vector after digested with Not Ⅰ and BamH Ⅰ, and ligated with T4 ligase. Results The constructed plasmid was checked by digestion with restrictive enzymes and DNA sequencing. Conclusion The plasmid of heavy chain of murine can be successfully constructed.
出处
《中国医学影像技术》
CSCD
北大核心
2009年第8期1355-1357,共3页
Chinese Journal of Medical Imaging Technology
基金
国家基础研究973基金(2006CB705707)
关键词
铁蛋白
重链
分子影像
报告基因
Ferritin
Heavy chain
Molecular imaging
Reporter gene
