电脉冲介导荧光素酶基因转移高效率的实验研究  

Electric pulse mediated high efficient luciferase gene transfer

作　　者：梅文瀚[1] 张兴黔[1] 钱关祥[1] 卢健[1] 丁隽[2] 

机构地区：[1]上海第二医科大学基础医学院生物化学教研室人类基因治疗研究中心,上海200025 [2]实验动物科学部

出　　处：《上海第二医科大学学报》2004年第8期624-626,634,共4页Acta Universitatis Medicinalis Secondae Shanghai

基　　金：国家自然科学基金(39970311);上海市科委重点项目基金(01JC14025);上海市教委重点学科基金(ZDXK20010)资助项目.

摘　　要：目的 研究电脉冲介导的基因转移效率及其最佳基因转移电脉冲参数。方法 用微量注射器将pcDNA3-Luc质粒100μg注射入BALB/小鼠的股四头肌,5 min内在注射部位给予不同参数的电脉冲刺激,然后进行荧光素酶活性测定和组织病理切片。结果 电脉冲可明显增加荧光素酶基因的表达,电脉冲组的荧光素酶活性(7.26±3.72)×107 RLU/mg蛋白是单纯注射组(1.38±1.27)×104 RLU/mg蛋白的5 262倍(P=0.017)。组织病理检查发现电脉冲刺激无严重的肌肉组织损伤。电压100 V/cm,波宽60 ms,脉冲次数10次和频率0.75 Hz是最佳的电脉冲参数。结论 在最佳的电脉冲参数条件下,电脉冲介导的基因转移可获得较高的基因表达。Objective To study gene transfer mediated by the electric pulse and optimize the parameters of electric pulse in vivo. Methods 100 μg plasmids of pcDNA3-Luc were injected into the quadriceps of BALB/c mice. Five minutes after DNA injection electric pulses with different parameters were given to the injection site. Two days after-ward, activity of luciferase was measured and the degree of muscle damage was analyzed by histological staining. Results The activity of luciferase in the electric pulse group [ (7. 26±3. 72) ×107 RLU/mg protein] was 5 262 folds higher than that in the simple injection group [ (1.38 ±1.27) ×104 RLU/mg protein] (P =0. 017). Histological analyses showed that in vivo electric pulse did not cause long-term severe muscle damage. When the electric pulse parameters were 100 V/cm,60 ms,10 pulses and 0.75 Hz, maximal gene expression was achieved. Conclusion Electri pulse,with optimal parameters, can increase gene expression obviously.

关 键 词：电脉冲 荧光素 酶基因 高效率 荧光素酶 基因转移 

分 类 号：R346[医药卫生—基础医学]