Team:Shanghai HS/Safety
General Lab Safety
Our lab at SIMM is classified as Biosafety Level 1, which means the lab uses microbes that pose little to no risk to healthy individuals. Hazardous chemicals and solutions are used at a minimal level and in well-ventilated areas such as fume hoods in the SIMM research lab. For instance, we do not use EtBr for gel electrophoresis. Instead, we use a safer nucleic acid stain called Seeing Safe DNA Dye. Nevertheless, all chemicals and solutions are still treated with all due respect, care, and caution. Our Laboratory belongs to the Shanghai Institute of Materia medica, Chinese Academy of Sciences. The bio-hazardous material has been used at a minimal level in the super clean bench, moreover, we choose the most environmentally friendly material that we can have access to the experiment. For example, we choose the agarose gel for electrophoresis (AGE) instead of polyacrylamide gel. Nonetheless, every reagent and microorganism that has been treated followed the standard lab criteria, and everyone responsible for experimenting has been trained by our instructors to correctly use the equipment and correctly follow the lab rules. No one eats or drinks in the lab for the entire process.
For most of the experiments that require plasmid reproduction, we use the common lab strain, the Escherichia coli (E. coli) K-12 DH5alpha. And all LB culture mediums that we used in cultivating the bacteria have been discarded correctly. The remaining bacterium is bleached then discarded. The other lab wastage like the pipette‘s spearhead is packed and sent to the professional disposal facility to ensure no contaminated materials are mismanaged. Moreover, for the PBS solution used in the ELISA test, we have the apparatus to wash those PBS out of the 96-well plate and discard those waste liquor into the specialized bottle which will be sent to the professional Waste liquid treatment plant. Two experienced research teachers Jianxia Ou and Rui Hu oversaw the training process and provide specific guides during experiments. To ensure the safety rules are tightly followed, we designed and agreed on a penalty system that charges anyone who breaks the rule a fine ranged from RMB 10-200. This penalty system successfully decreases the rate of inappropriate and false practice and the money collected is used as funds for the project.
Fig.1. Picture of our laboratory where our iGEM experiments are conducted.
Project Bio-safety
Synthetic Pseudo-virus
Instead of using actual viruses, which are dangerous and restricted without exception, we used synthetic pseudo-virus. The purpose of using it is to simply make an identification of the inhibitory activity of antiCovid-19 monoclonal antibody against the epidemic mutant B.1.1.7
The pseudo-typed B.1.1.7, B.1.351, and B.1.617 were constructed by co-transfection of two plasmids, one expressing Env-defective HIV-1 with luciferase reporter (pNL4-3.luc.RE)[1] and the other expressing the full-length S-protein of B.1.1.7, B.1.351, or B.1.617 into HEK293T cells.
Reference
1.Xia, S. et al. Inhibition of SARS-CoV-2 (previously 2019-nCoV) infection by a highly potent pan-coronavirus fusion inhibitor targeting its spike protein that harbors a high capacity to mediate membrane fusion. Cell Res 30, 343-355, doi:10.1038/s41422-020-0305-x (2020).
2.http://www.simm.cas.cn/web/jgsz/yjbm/index_83071.html ?json=http://sourcedb.simm.cas.cn/zw/gb2020/yjzz/202008/t20200825_5676100.json
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