摘要
目的:构建pLEGFP-mIFNγ真核表达质粒,以获得带有绿色荧光蛋白的γ干扰素融合蛋白。方法:以克隆质粒pcDNA3.1-mIFNγ为模板,用PCR方法扩增mIFNγDNA片段,利用pEGFP-C1质粒载体构建pLEGFP-mIFNγ重组质粒,用酶切电泳验证重组质粒的正确性。脂质体转染包装细胞PA317,利用PA317病毒上清转染神经干细胞。结果:PCR结果显示扩增片断大小约500 bp,与预期相同。重组质粒酶切后显示其大小约7 400 bp。pLEGFP-mIFNγ成功转染神经干细胞。结论:pLEGFP-mIFNγ质粒构建成功,能够有效转染神经干细胞,并进行示踪。
Objective To construct a recombinant plasmid pLEGFP-mIFNγ/ for developing the genetical therapy of tumor by stem cells. Methods PCR was used to amplify the mIFNγ/gene from plasmid pcDNA3.1-mIFNγ/in which the mIFNγ/ was cloned. A new plasmid, pLEGFP-mIFNγ/, was then constructed by inserting the amplified mIFNγ/ gene into pLEGFP-C1. Restriction analysis and sequencing were used to confirm the structure of pLEGFP-mIFNγ/. Neural stem cells were transfected by supernatant of PA317. Results A DNA fragment in size of 500 bp was amplified. Restriction analysis showed that the amplified gene was inserted into pLEGFP-C1 correctly. Conclusion The plasmid pLEGFP-mIFNγ/ is constructed successfully, and it can transfect neural stem cells.
出处
《吉林大学学报（医学版）》
CAS
CSCD
北大核心
2006年第4期593-595,F0002,共4页
Journal of Jilin University：Medicine Edition
基金
吉林省科技厅科技发展计划项目资助课题(20030542-1)
吉林省长春市科委资助课题(04-07SF030)
