Cloning Expression and Functional Verification of recF and recO Gene from Deinococcus radiodurans R1 in Escherichia coli / 环境与健康杂志

ABSTRACT

Objective To construct expressing recombinant of recF and recO from Deinococcus radiodurans R1and express the target protein in E.coli BL21(DE)3.Methods recF and recO gene were amplified by PCR from genomic DNA of Deinococcus radiodurans R1 and inserted into expression plasmid vector pET30b(+) to construct pET30b(+)-recF and pET30b(+)-recO.The recombinant plasmids were transformed into E.coli BL21(DE)3 and the recombinant proteins were expressed by the isopropyl-?-D-thiogalactopyranoside(IPTG) and were analyzed with SDS-PAGE.The changes in the survival rate of bacteria in each group before-and after UV-irradiation were calculated.Results The recombinant plasmid pET30b(+)-recO and pET30b(+)-recF was obtained and the recombinant protein could be highly expressed in E.coli BL21(DE)3.Conclusion This study has provided a foundation for further studies and applications of the recombinant RecF and RecO,and initial detection shows that recO and recF gene can increase the resistant ability of E.coli.