超声波介导质粒pET28a转化枯草芽胞杆菌的研究

Ultrasound Media-Induced Plasmid pET28a to Transform Bacillus subtilis

揭示超声波介导下质粒DNA转化枯草芽胞杆菌发生的生物学机制,以便于枯草芽胞杆菌基因工程菌株的工业化应用。建立超声波介导质粒pET28a转化枯草芽胞杆菌的一种方法,采用扫描电镜结合理化分析检测技术研究超声波处理前后细胞的变化。结果显示,一定条件的超声(40~100 W)处理后的质粒DNA可以转化枯草芽胞杆菌,处理后的细胞在电镜下表面凹凸不平,出现褶皱现象;而且胞内蛋白质、磷脂及碱性磷酸酶(AP)大量释放至胞外。结果表明,超声波介导的DNA转化是细胞生理响应与超声波共同作用的结果。

The biological mechanism happened in the transformation of Bacillus subtilis under media-induced plasmid DNA with ultrasound was revealed, in order to apply B. subtiliz genetic engineering strain industrially. A general method to media-induce plasmid pET28a and to transform B. subtilis ultrasound was established, and studied the changes of cells adopting electronic microscope combined with physico-chemical analyses technologies before and after ultrasound treatment. The results showed that plasmid DNA after treated with a certain condition of ultrasound （40 - 100 W） could transform B. subtilis; after the treatment the surface of cells were ravelled and rimpled under the elec- tron microscope; furthermore, a large amount of intracellular proteins, phospholipids, alkaline phosphatase and （AP） release to outside the cell. Therefore, media-induced DNA by ultrasound to transform is the results of combined action of cell physiological responses and ultrasound.