Expression And Characterization Of Aldehyde Ketone Reductase AKR7A3 In Liver Cells

AKR7A3 is an aldosterone reductase(AKR),which is one of the NADPH dependent oxidation reductase superfamilies.AKR7A3 is involved in many extremely important physiological activities in vivo,such as hormone synthesis,drug delivery,inflammatory reaction and the process of detoxification.AKR7A3 is expressed in the process of carcinogenesis in liver,kidney and uterus.As aflatoxin detoxification enzyme is widely known.Existing studies can prove that the development of the enzyme and liver cancer is inextricably linked.Taking AKR7A3 as the research object of this experiment,establish AKR7A3-pET-15 b expression vector,using e.coli BL21 aldehyde ketone reductase AKR7A3 prokaryotic expression,to express the corresponding optimized conditions,and determination of prokaryotic expression of proteins related to the activity.In vivo experiments,a multi-antibody model of AKR7A3 mouse was established to obtain polyclonal antibodies.Hl-7702 and HepG2 of hepatocellular carcinoma cells were used to investigate the correlation between the enzyme and liver cancer from the perspective of proteins and nucleic acid molecules in the cells using the techniques of ELISA and RT-PCR.The experimental results showed that a large number of highly purified human AKR7A3 with biological activity was obtaiued,the optimized condition results were 16 degrees Celsius,110 rpm of rotational speed,20 hours of time,and 1.00 mM/L of IPTG.The immune living animals get the antibodies.The results of the expression of this enzyme in both cells were 2.1 times higher than that of liver cancer cells at the protein level and 2.84 times higher than that of the liver cancer cells at the nucleic acid level.