低频低强度超声介导质粒转入大肠杆菌的实验研究  

Experimental Study of Low-Frequency and Low-Intensity Ultrasound-Mediated Plasmid Transferring into E.coli

作　　者：郑慧敏[1] 江和逊 苏杭[1] 董宇[1] 李岱容[2] 杜永洪[1] 

机构地区：[1]重庆医科大学生物医学工程学院,省部共建国家重点实验室培育基地-重庆市超声医学工程重点实验室,重庆市生物医学工程学重点实验室,重庆市400016 [2]重庆医科大学附属第一医院

出　　处：《中国超声医学杂志》2016年第9期837-839,共3页Chinese Journal of Ultrasound in Medicine

基　　金：国家自然科学基金项目(No.81101216;81127901);国家"973"计划项目(No.2011CB707900);重庆医科大学资助项目(No.NSFYY201529)

摘　　要：目的探讨不同因素对低频低强度超声(LFLIU)介导质粒转入大肠杆菌的效率影响。方法大肠杆菌和质粒DNA混合液用42kHz,0.13~0.72 W/cm^2超声辐照后,在氨苄霉素琼脂培养板上进行菌落计数,计算质粒转化效率。同时,检测不同强度超声对大肠杆菌存活率和质粒完整性的影响。结果 (1)大肠杆菌和质粒混合液经LFLIU辐照后,能成功在氨苄霉素琼脂培养板上筛选出携带抗性的菌株,质粒转化效率与超声辐照剂量和CaCl_2浓度有关。(2)流式细胞仪检测结果表明,0.13~0.72 W/cm^2的超声辐照对大肠杆菌的活力无显著影响。(3)电泳检测结果显示,不同强度超声辐照质粒,随着超声强度的增加,导致质粒构型改变,但不影响其结构的完整性。结论 LFLIU能够介导质粒转入大肠杆菌,可为LFLIU将质粒和基因转入其他特殊细菌提供实验参考。Objective To study the influence of different factors on efficiency of low-frequency and low-intensity ultrasound mediated plasmid transferring into E. coll. Methods A mixture of E. coli and plasmid were exposed to 42 kHz ultrasound with the intensity at 0.13-0.72 W/cm^2, respectively. Next, the bacterium liquid was plated onto ampicillin agar media for colony count, and transformation efficiency was calculated. Meanwhile, the survival rate of E. coli and plasmid integrity were detected at the intensity of ultrasonic aforementioned. Results （1）Positive olonies were selected on the LB agarose plate containing ampicillin after ultrasound exposed. （2）The result of flow cytometry showed the different intensity aforementioned had no significant effect on the survival rate of the bacteria. （3）The electrophoresis detection showed the configurations of plasmid were ehanged by higher intensity of ultrasonic, but the integrity were not changed. Conclusions LFLIU can mediate plasmid transferring into E. coli efficiently. The results can provide experimental reference for LFLIU-mediated plasmid and DNA transformation into other special baeteria.

关 键 词：低频低强度超声 大肠杆菌 质粒 转化效率 

分 类 号：R445.1[医药卫生—影像医学与核医学]