Thermophilic bacteria and viruses represent novel sources of genetic materials and enzymes with great potential for use in industry and biotechnology. In this study, GVE2, a virulent tailed Siphoviridae bacteriophage infecting deep-sea thermophilic Geobacillus sp. E263, was characterized. The bacteriophage contained a 40,863-bp linear double-stranded genomic deoxyribonucleic acid (DNA) with 62 presumptive open reading frames (ORFs). A viral DNA microarray was developed to monitor the viral gene transcription program. Microarray analysis indicated that 74.2% of the presumptive ORFs were expressed. The structural proteins of purified GVE2 virions were identified by mass spectrometric analysis. The purified virions contained six protein bands. Of the newly retrieved proteins, VP371 was further characterized. The immuno-electron microscopy indicated that the VP371 protein was a component of the viral capsid. Transcriptional analyses and proteomic characterization of GVE2 would be helpful to understand the complex host-virus interaction during virus infection.