不同方式递送质粒DNA诱导体内表达效果的实验研究  

Plasmid DNA induces in vivo expression of differently delivered gene

作　　者：张亮[1] 李潇潇[2] 韩刚[1,3] 董金凯[1,3] 阎瑾琦[1] 肖毅[1,3] 于继云[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850 [2]北京积水潭医院检验科,北京100035 [3]解放军总医院泌尿外科,北京100853

出　　处：《军医进修学院学报》2010年第1期48-49,60,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：国家"863"基金项目(2006AA02A237; 2007AA02Z451);国家自然科学基金项目 (30772002)~~

摘　　要：目的比较不同途径递送质粒DNA至Balb／c小鼠体内所诱导的报告基因表达效果。方法将编码荧光素酶（1uc3蛋白的重组质粒（pGL-3-CMV）或空载体质粒pGL-3-basic以肌肉注射或电脉冲方法将10μg或100μg上述质粒注入小鼠股四头肌，基因枪法以三枪接种质粒于小鼠腹部（2μg质粒DNA／4．5Mpa／枪）。于质粒注入后24～144h，检测小鼠体内的荧光素酶活性。结果肌肉注射10μg的小鼠未检测到荧光素酶的表达；肌肉注射100μg、电脉冲法递送10μg和100μg质粒的小鼠，接种后48h体内荧光素酶活性达到峰值，递送100μg的小鼠体内表达量明显高于10μg的小鼠。基因枪免疫小鼠在接种24h达到峰值。结论电脉冲和基因枪介导的基因递送方式基因表达水平远远高于传统注射方法，是基因疫苗免疫递送的可靠方法。Objective To explore the differently delivered luciferase gene expressions in Balb/c mice. Methods Balb/c mice received intramuscular injection of recombinant luciferase-expressing plasmid, pGL-3-CMV or its mock plasmid, pGL-3-base, and electric, respectively. 10 μg or 100 μg of the above plasmids was intramuscularly injected or via electric pulse into musculus quadriceps fexoris. The plasmids were intradermally injected with a gene gun through 3 bullets, 2 IX g of plasmids and 4.5Mpa for each shoot. Luciferase activities were monitored for 24-144 hours after the gene was delivered into mice by bioluminescence imaging in vivo. Results The luciferase activity was not detectable in the mice after intramuscular injection of 10μ g pGL-3-CMV, and reached its peak 48 hours after injection of 100 μg pGL-3-CMV, and was detectable in mice injected with 10μ g or 100μg pGL-3-CMV using a gene gun. After 24 hours, the luciferase activity reached its peak. Conclusion The exogenous gene expression level induced by injection with electric pulse and gene gun is higher than that induced by traditional intramuscular injection. It is reliable to use electric pulse and gene gun to deliver gene.

关 键 词：基因递送 质粒 DNA 荧光素酶基因 生物弹道技术 

分 类 号：R392.7[医药卫生—免疫学] R392-33[医药卫生—基础医学]