超声波介导的微生物细胞转化

Ultrasonic-mediated DNA transformation of microbial cells

摘要:

随着分子生物学的发展, 微生物遗传改造越来越广泛地应用在微生物育种、临床医学、环境保护等方面。其中, DNA转化技术经常是高效遗传改造的瓶颈之一。应用超声波将目的基因导入微生物细胞的技术具有原位、多尺度、活体、高通量、低成本等优点, 因此发展较为迅速。其原理是超声波可以通过声学气穴现象产生一系列的非热能效应, 而声学气穴微泡可产生短暂的细胞膜透化作用。本文综述了超声波转化的基本原理及其在微生物细胞转化中的发展现状, 并结合本实验室应用超声波转化法转化革兰氏阳性菌等研究进展, 分析了其特色、优势及现存挑战。

Abstract:

With the development of molecular biology, genetic engineering of microorganisms has been increasingly applied in microbial breeding, clinical medicine and environmental bioremediation. However delivery of DNA into the cell can be one bottleneck in efficient genetic engineering. Ultrasound-mediated DNA delivery into microbial cells, with a number of advantages that include in situ operation, spatial scalability, non-invasiveness, high-throughput and low consumable costs, has been under rapid development. Ultrasound can produce a variety of nonthermal effects via acoustic cavitation, and the cavitation bubbles generated during this process can cause transient cell membrane permeability. In this review the basic principles of the ultrasound-based DNA delivery were discussed and its current applications in prokaryotes were summarized. Furthermore, we discussed the advantages and challenges of the technique, using the transformation of Gram-positive bacteria in our laboratory as an example.