| Modern fermentation engineering and technology is considered to be an important technology of the 21 st century. It’s one of the symbols of the new technology revolution. The countries all over the world has paid attention to it. Modern fermentation engineering has great potential to provide the most promising solutions to solve the problem :environment, resource, population, energy and food crisis and pressure. And it’s closely related to many fields such as the chemical industry, medicine, food, energy, environmental protection and agriculture. There is great economic benefits to develop related technologies of modern fermentation engineering and technology.Fermentation engineering is a new technology, refers to the use of modern engineering technologyand the specific function of microorganisms to product useful products for human being or directly apply microbes in the process of industrial production. Fermentation technology has had a great development through decades development,but there are still many factors constraint the efficiency of production.As two pillars of modern fermentation engineering, recombinant DNA technology and biological reactor, are two big bottleneck to be broken through. In modern fermentation engineering, many times you need to delivery the modified genes into microbial, and then enlarge the culture scale in order to produce plenty of metabolites.. However in the process, there are some defects as follows: a, the traditional transformation method is relatively complicated and time-consuming; b, the process of cryopreservation can lead to vigor recession and you need to reculure the strain in order to recovery the strain; c, in large-scale industrial production, the process of enlarged culture is time-consume and cumbersome to operate.We hope to have a new transformation technique which can directly delivery the gene into microbial in the bioreactor efficiently and on a large scale.in this way we can quickly and efficiently implement transformation and avoid the tedious process of enlarge cultivate. The application of ultrasonic to delivery genes into microbial cell as a new and immaturity transformation technology has entered our field of vision.The basic principle of Ultrasound-mediated DNA delivery :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. 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. The development and utilization of the new technology can help solve the above problems, improve the production efficiency of microbial fermentation.This research is undertaken under this background, as a basic reasearch,it has important significance to promote the development of Molecular biology and fermentation engineering etc.On the basis of the previous studies, this study is committed to further simplify and improve the ultrasonic technology, to solve the above problems in fermentation engineering. The obtained results are as follows:In this reserch, we take E. coli BL21(DE3) as the vector, successfully delivery the plasmid p ET-20b(+) into the cell under the optimized conditions. Measured the optimum transformation conditions of the simplified ultrasonic technology under different standards, the highest conversion rate is 42 CFU/?g plasmid when the OD600 is 9, plasmid concentration is 0.5 ng/?l, ultrasonic power is 30 w, conversion time is 180 s. |
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