抄録
Deinococcus radiodurans ddrO gene have played an important role in the process to withstand radiation and oxygen free radical damage. PCR amplification ddrO gene and cloned ddrO gene into pGEM-T easy Vector . Shuttle express vector pRADZ3(+)ddrO is constructed and identified by using restriction enzyme SpeI and NdeI double digestion for recombinant T(+)ddrO and shuttle vector pRADZ3 respectively,Connect ddrO fragment and big fragment of shuttle vector pRADZ3 through T4 DNA ligase. Identification by PCR and digestion, confirmed that recombination shuttle vector pRADZ3(+)ddrO constructs successfully. Transforms the recombination shuttle vector to E.coli DH5α competence cell, let it to express the DdrO protein under the normal culture condition (not to need Inducer),and through Western blot confirmed that the ddrO gene can express stably in DH5α. Compare with DH5α which transformed empty vector pRADZ3 ,and observing the change of this two kind of E.coli under the ultraviolet exposure and the H2O2 oxidation pressure survival ratio, studies the DdrO protein anti-ultraviolet radiation and the against oxidized damage function in E. the coli DH5α. The experimental result demonstrate that ddrO gene express in E. coli be able to strengthen it ability to bears the ultraviolet radiation and the strong oxidant H2O2.
