Overview
Escherichia coli, as an important model industrial microorganism , has extensive applications in the fields of pharmaceuticals, chemicals, agriculture, and experiments.
We adopt various new strategies and technologies for metabo sign, construct, and optimize E. coli chemical cell factories. During this process, safety issues cannot be ignored. We have analyzed and considered the entire project and listed the following contents.
Experimental Material Safety
Our project is to mine the to bear the saline-alkaline environment of universal genetic parts which can enhance the ability of engineering bacteria (such as Escherichia coli) in salt-resistant bacteria.
Six strains with good saline-alkali resistance were collected from dung beetle intestines, Populus euphratica xylem, uranium mineral, saline lake, Aidin Lake and saline-alkali land. They are: Brevibacterium casei G20, Bacillus haynesii P19, luteus R17, bluephagenesis TD01, Halomonas Campaniensis LS21,Enterobacter cloacae RS35,Bacillus haynesii P19, Micrococcus luteus R17, Enterobacter cloacae RS35 is a safety class 1 microorganism, Brevibacterium casei G20 is a safety level 2 microorganism, as for Halomonas bluephagenesis TD01 and Halomonas Campaniensis LS21, we have submitted the relevant check-in form. We mainly carry out the stage of component excavation and testing. In the laboratory stage, we will strictly control the strains used for excavation and the recombinant engineering bacteria used for testing, and will not be brought outside the laboratory.
When we dig up the target gene, we will transfer them into E.coli DH5 α, E.coli S17-1, E.coli BL21 (DE3) and Bacillus subtilis for testing. E.coli DH5 α, E.coli S17-1, E.coli BL21 (DE3), Bacillus subtilis are all safety level 1 microbes. The test strains we use are not dangerous and we will kill them strictly after the test and will not take them out of the laboratory.
In the next few years, if our project fully developed, these recombinant engineering bacteria may be put into factory fermentors, test fields or be used for oil field production. However, before putting into commercial use, we strictly abide by theEnvironmental Protection Law, Biosafety Law, Agricultural Law and so on to evaluate them. For example, these strains implanted with genetic parts will have better saline-alkali resistance, so will they be more competitive than other bacteria in the environment, thus affecting the growth of other bacteria in the environment, and if so, whether they will cause damage? If it causes damage, we will refuse to use these strains in an open environment.
Experimental process safety
During the experimental process, there are two main dangers.
1. Hazards in terms of reagents. The highest pH of the culture medium we used reached pH=14, which may result in careless operation during the experiment, causing the medium to overturn and come into direct contact with the experimenter's skin and eyes. For this purpose, we have equipped each experimental personnel with experimental clothing, professional rubber gloves, and goggles. The laboratory is also equipped with emergency shower devices to protect the safety of experimental personnel as much as possible.
2. Hazards related to instruments. We will use high-pressure sterilization pots and ultra clean workbenches during the experimental process. During use, the high-temperature steam generated by the high-pressure sterilization pot may cause burns to the experimental personnel; The ultraviolet radiation emitted by the ultra clean workbench can directly come into contact with the experimenter's eyes and cause eye burns.
To prevent this situation from happening, we provided training to the members of the experimental group before starting the experiment of this project, allowing them to learn about experimental operations and the use of instruments, and equip them with the ability to respond to unexpected situations. At the same time, we have equipped professional teachers to regularly check the use of experimental equipment, and try to avoid injuries to experimental personnel caused by damage to experimental equipment.
