Team:BUCT-China/Design
Design
Engineering principles:
We have found an oxidase gene from a Marine bacteria, which can degrade polymers like polyolefin. After testing, we found that it has a strong degradation ability, and the degradation mechanism is different from the previous research. Therefore, we registered it as parts, hoping that it could be used for your study.
Design iterations:
We commissioned the company to synthesize the plasmid main chain and the gene of this oxidase, and then used the enzyme to connect oxidase gene to the plasmid.
Experimental steps:
The plasmid was introduced into E.coli.
The bacteria were cultured in polyethylene with inducer.
Oxidase expression test
Measurment of the enzyme activity：
Oxidase activity was measured with ABTS as substrate, and the absorbance change was detected at 420nm with a spectrophotometer for 3min.
Protein expression：
Protein levels were detected by polyacrylamide gelelectrophoresis and oxidase gene expression was determined by comparing with the standard.