| Background and objective: Deinococcus radiodurans(DR)is one of the organisms found on earth with extreme resistance to ionizing radiation.PprI acts as a general switching factor of radiation in DR PprM is a novel regulatory protein of PprI dependent radiation response.The aim of this study was to construct a drosophila model of stable genetic expression of anti-radiation gene pprM/pprI.This research that involved in the effects of pprM/pprIon the transgenic drosophila melanogaster,may inspireus with new ideas for the treatment of radiation damage.In addition,the results of this study provide the theoretical basis for monitoring the leakage dose of nuclear accident,mechanism of radiation resistance and human radiation protection.Methods:1.PprM gene was amplifiedby PCR using pGEX-6p-1-pprM pdsred1-n1-flag-pprI as template,and pattb-UAST-HA-pprM and pattb-UAST-HA-pprI vectors were constructed.2.pattb-UAST-HA-pprM and pattb-UAST-HA-pprI vectors were injected into yw type drosophila embryo by microinjection of drosophila embryo.Construction of transgenic Drosophila model.3.Total superoxide dismutase(SOD)activity,catalase(CAT)activity and TBA microplate method were used to detect malondialdehyde(MDA)and reduced glutathione(GSH).The oxidative damage ofDrosophila was evaluated before and after irradiation.Results:1.Sequencing Identification and Blast comparative analysis showed that pattb-UAST-HA-pprM and pattb-UAST-HA-pprI weresuccessfully constructed.2.Stable genetic pprI/pprM transgenic Drosophila were obtained by microinjection and screening,and pprM/ pprI was identified by phenotype on chromosome 2 and chromosome 3 of Drosophila,respectively.3.The oxidative resistance and radiation resistance of transgenic pprI Drosophila were improvedConclusion: The protein overexpression vector of PprM/PprI was successfully constructed,and four strains of transgenic Drosophila were obtained through microinjection and screening,which verified the anti-radiation ability of transgenic Drosophila?... |
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