A novel Pseudoalteromonas marina bacteriophage, PH101, specifically infecting Pseudoalteromonas BH101 was isolated from the water sample of the Yellow Sea of China using the agar overlay method. 16S rDNA sequence identification was used to identify the host bacteria. Efficiency of infection, multiplicity of infection value, morphological characterization, one-step growth curve, and host range of the bacteriophage were determined. Purified PH101 genomic DNA was extracted and its genome was completely sequenced and analyzed. The phage morphology showed that PH101 belongs to the Myoviridae family with a head of 60 nm in diameter and a tail of 40 nm with a tail fiber of 10-20 nm. Microbiological characterization demonstrated that phage PH101 is stable at a wide range of temperatures (0-70 °C) and showed acid and alkaline resistance (pH 3-12). The one-step growth curve showed a latent period of about 20 min, a rise period of 20 min, and a burst size of about 31.6 virions. The genome sequencing and bioinformatic analysis shows that phage PH101 was a novel bacteriophage which was found to consist of a linear, double-stranded 131,903-bp DNA molecule with a GC content of 37.36 % and 228 putative open reading frames without RNA, which were classified into seven functional groups, including phage structure, adsorption, packaging, gene transfer protease, terminase, DNA binding, and regulation.