Kant Rishi, Bhatt Geeta, Patel Vinay Kumar, Ganguli Anurup, Singh Deepak, Nayak Monalisha, Mishra Keerti, Gupta Ankur, Gangopadhyay Keshab, Gangopadhyay Shubhra, Ramanathan Gurunath, Bhattacharya Shantanu
Microsystems Fabrication Laboratory, Department of Mechanical Engineering and Department of Chemistry, Indian Institute of Technology Kanpur, Kanpur 208016, Uttar Pradesh, India.
Department of Electrical and Computer Engineering, University of Missouri Columbia, Columbia, Missouri 65211, United States.
ACS Omega. 2019 May 15;4(5):8512-8521. doi: 10.1021/acsomega.9b00202. eCollection 2019 May 31.
We report a simple device that generates synchronized mechanical and electrical pressure waves for carrying out bacterial transformation. The mechanical pressure waves are produced by igniting a confined nanoenergetic composite material that provides ultrahigh pressure. Further, this device has an arrangement through which a synchronized electric field (of a time-varying nature) is initiated at a delay of ≈85 μs at the full width half-maxima point of the pressure pulse. The pressure waves so generated are incident to a thin aluminum-polydimethylsiloxane membrane that partitions the ignition chamber from the column of the mixture containing bacterial cells ( BL21) and 4 kb transforming DNA. A combination of mechanical and electrical pressure pulse created through the above arrangement ensures that the transforming DNA transports across the cell membrane into the cell, leading to a transformation event. This unique device has been successfully operated for efficient gene (∼4 kb) transfer into cells. The transformation efficacy of this device is found comparable to the other standard methods and protocols for carrying out the transformation.
我们报道了一种简单的装置,该装置可产生同步的机械压力波和电压力波以进行细菌转化。机械压力波通过点燃一种受限的纳米含能复合材料产生,该材料可提供超高压。此外,该装置有一种设置,通过该设置,在压力脉冲的半高宽点处延迟约85微秒启动一个同步电场(随时间变化)。如此产生的压力波入射到一个薄的铝-聚二甲基硅氧烷膜上,该膜将点火室与含有细菌细胞(BL21)和4 kb转化DNA的混合物柱分隔开。通过上述设置产生的机械和电压力脉冲的组合确保转化DNA穿过细胞膜进入细胞,从而导致转化事件。这种独特的装置已成功运行,可将高效基因(约4 kb)转移到细胞中。发现该装置的转化效率与其他用于进行转化的标准方法和方案相当。