Objective：Isolate and identify a new strain of lytic phage and study its biological characteristics，so as to provide an alternative treatment for clinical infection caused by multi-drug resistant bacterial in the future. Methods：The new phage named vB_EcoM_RZ was isolated and purified by double layer agar method. The morphology of the new phage was observed by electron microscope. The incubation period and burst size of the new phage were determined by one-step growth curve，the genomic characteristics and encoded proteins of phage vB_EcoM_RZ were studied through the whole genome sequencing and mass spectrometry analysis combined with bioinformatics analysis. Results：A new type of Escherichia coli virulent bacteriophage was successfully isolated；biological experiments showed that it had fast lytic speed，wide host range，and good temperature stability；genomics studies showed that it did not encode genes related to toxins or other virulence factors；43 known proteins and some hypothetical proteins with unknown functions were isolated and identified by mass spectrometry. Phylogenetic analysis showed that it was a new type of bacteriophage. Conclusion：A new type of Escherichia coli virulent bacteriophage named vB_EcoM_RZ has been isolated and identified. It has excellent properties and the potential to be further used in the treatment of infection caused by drug-resistant bacteria.