Abstract: Resuscitation-promoting factor（Rpf） is a type of cytokine involved in the resuscitation of dormant cells and bacterial growth. Rhodococcus erythropolis KB1 Rpf and petroleum-contaminated soil were added to the bacterial enrichment medium containing crude oil to observe the effect of Rpf on the crude oil degradation ability and enrichment of culturable bacterial diversity of indigenous microorganisms in contaminated soil. The oil degradation rates in the groups with different recombinant Rpf were increased from 30.42% to 36.51% and 37.93%, respectively. The recombinant Rpf also increased the diversities of the cultivable oil-degrading bacteria. In the control group without Rpf, nine bacterial phyla were determined. Proteobacteria was the dominant bacterial phylum, with a relative abundance of 94.55%. 82 bacterial genera were founded. The dominant bacterial genera were Acinetobacter, Pseudoxanthomonas and Achromobacter, with relative abundances of 55.32%, 6.94% and 6.32%. In the groups with recombinant Rpf, the bacterial phyla were increased to 13 and 16 respectively. The Proteobacteria was the dominant bacterial phylum, with the relative abundances of 92.64% and 95.21%. And in the groups containing the recombinant protein 99 and 122 bacterial genera were founded. The abundances of Acinetobacter were decreased to 24.96% and 33.25%, but abundances of Stenotrophomonas increased from 1.85% to 23.20% and 32.26%. Abundances of Ochrobactrum increased from 0.94% to 6.97% and 1.15% respectively. Six oil degrading bacteria were isolated from the oil contaminated soils in the groups containing the recombinant Rpf. In this study, we clarified the effect of Rpf protein on the diversity of soil petroleum-degrading bacteria and isolated efficient petroleum-degrading bacteria, which provided a new method and idea for the bioremediation of petroleum-contaminated soils.