Functions Of Rhodaneses In Musca Domestica And Macrobrachium Nipponense

House fly (Musca domestica) is an insect of Diptera. It widely distribute all over theworld. Macrobrachium nipponense is an aquatic arthropods. It widely distributes in the riverand lake in China, Japan, southeast Asia and far east of Russia. This study used the housefliesand Japan spermatogenesis as materials, researched and compared the function of Rhodanese.This study contains4parts:(1) A sequence of rhodanese obtained in the previous study was used as the querysequence.It was used to find potential sequence of rhodanese in1transcriptome of Muscadomestica and2transcriptome of Macrobrachium nipponense.We obtained5new sequencesin transcriptome of Musca domestica and3new sequence in transcriptome of Macrobrachiumnipponense. These sequences were named Md-Rho1-5and Mn-Rho1-3.(2)6vector of prokaryotic expression were constructed successfully.4kinds of solublerecombinant protein rMd-Rho1, rMd-Rho4, rMn-Rho1and rMn-Rho2were obtained andeach has rhodanese activity. Also2kinds of recombinant protein inclusion bodies rMd-Rho2and rMd-Rho3were obtained, and after refolding, these2recombinant proteins also showedrhodanese activity.(3) GST pull down assay was used to find the protein interacting with rhodaneses.Weobtained a protein with molecular weight of about80kD, but failed to identify it with massspectrometry.(4) Real-time quantitative PCR technique was used for the study of gene expressionpatterns of Md-Rho3, Md-Rho4，Mn-Rho1and Mn-Rho2. Md-Rho3, Mn-Rho1, Mn-Rho2upregulated after bacterial challenge, while Md-Rho2was downregulated after the sametreatment. The result of gene expression in different tissue showed Md-Rho3mainlyexpressed in the haemocyto and Md-Rho4mainly expressed in the intestine, Mn-Rho1mainlyexpressed in the gills and liver and Mn-Rho2mainly expressed in the gill, revealing thedifferent roles of rhodaneses in immunity.