The Preliminary Reasearch Of Anti-oxidant Function Of Rhodanase From Macrobrachium Nipponense

Macrobrachium nipponense can be regarded as important fresh water shrimps.Because it was easily suffered from various infections,which is often caused enormous economic loss.The research of the immunologic mechanism with antioxidant has great significance.It was studied that rhodanase have the function with antioxidant in Azotobacter and mammal cells.We once studied a M.nipponense rhodanase sequence,and this sequence was used to find potential sequence of from transcriptome of M.nipponense,and 4 sequence of rhodanase has been discovered,which were named MnRHOD2、MnRHOD3、MnRHOD4、Mn RHOD5.The inducible expression of MnRHOD2-MnRHOD5 of rhodanese of M.nipponese has been successfully realized through the research,and the activity of corresponding fusion proteins has been also checked,which shows the result that all the four inducible expression proteins possess enzyme activity characteristics.Furthermore,the expression modes of the four proteins are not the same.The genes of MnRHOD2 MnRHOD3 and MnRHOD5 are relatively the highest in the part of hepatopancreas while MnRHOD4 are the highest in the part of gills.The three genes of MnRHOD2 MnRHOD4 and MnRHOD5 show upward trend with the stimulation of Aeromonas veronii,while MnRHOD3 show the downward trend.To prove the relationship between the gene of rhodanese of M.nipponense and the oxidant stress,MnRHOD2 are knocked down through the method of RNAi.With knocking down the MnRHOD2,the level of ROS and MDA has been raised.To further prove whether the rhodanese of M.nipponese have antioxidation or not,when PMS emerge oxidative stess,the numbers of survival strain of E.coli after active site mutation is remarkably less than those before active site mutation.These preliminarily proved that the rhodanese have anti-oxidant stress.