电激和普鲁卡因增强DNA导入哺乳动物活体效率  

Electric Shock and Procaine Enhanced DNA Transfection Efficiency in Mammalian

作　　者：李传宝[1,2] 华甜[2] 金海明 杜宏武[1,2] 

机构地区：[1]北京科技大学生物工程与传感技术研究中心 [2]北京科技大学化学与生物工程学院生物技术系,北京100083

出　　处：《生物技术》2011年第6期49-52,共4页Biotechnology

基　　金：国家自然基金项目("新的hMGF相互作用蛋白筛选及其介导肌肉再生的分子机制";No.30800540)资助

摘　　要：目的:探究电激和普鲁卡因对DNA导入哺乳动物活体效率的影响。方法:向小鼠的股四头肌注射20μgβ-gal质粒,分别采用以下方法:直接注射;先注射100μL 0.25%的普鲁卡因,24 h后注射质粒;注射质粒后进行经过前期优化后的电激程序进行加强(500 V,4个脉冲)。注射后的第2 d、1w和2w后进行x-gal染色检测。结果:三种导入方法均可将外源质粒导入肌肉组织中并进行表达。电激和普鲁卡因可以显著增强肌肉对外源基因的吸收和表达,尤其是优化后的电激程序,其可以使肌肉长期稳定高效的表达外源导入基因。结论:普鲁卡因是常用的增强DNA导入效率的促进剂,而合适的电激可以更为显著地增强DNA导入效率,均是基因治疗中提高基因导入效率的良好候选方案。Objective:To explore whether electric shock and procaine can enhance DNA transfection efficiency in mammalian.Method:Mice were randomly divided into 3 groups.The quadriceps from group 1 were injected 20 μg β-gal plasmid directly;Group 2 were firstly injected 100 μL of 0.25% procaine and 24 hours later 20 μg β-gal plasmid was injected to the same location;For the group 3,20 μg β-gal plasmid was injected into quadriceps then followed by electric shock 4 times(500 V),which was pre-optimized.At 2 days,1 week and 2 weeks after injection,the β-gal expression was detected by x-gal staining.Result:It was found that all the three methods could transfect β-gal plasmid into mouse muscle tissue and β-gal was expressed well.Electric shock and procaine both can facilitate the mouse muscle to absorp and express foreign gene,especially electric shock can make β-gal express in muscle for long-term,more stably and efficiently.Conclusion:Electric shock enhanced the efficiency of DNA transfection into muscle in vivo.This is a good strategy to improve the efficiency of gene transfer in gene therapy.

关 键 词：电激 基因治疗 DNA导入 普鲁卡因 人半乳糖苷酶 

分 类 号：Q81[生物学—生物工程]