Previously, we demonstrated the presence of a second copy of LPS myristoyl transferase in enterohemorrhagic Escherichia coli O157:H7, an important zoonotic diarrheagenic food-borne pathogen; the pO157-encoded ecf (an eae-conserved fragment) and the chromosomally-encoded lpxM (also referred to as msbM) genes. Although both genes share the same function as an LPS myristoyl transferase, the pO157-encoded ecf is thermoregulated via an intrinsically curved DNA while the chromosomal lpxM is regulated by the PhoP/Q two component regulatory system. However, it is unclear why E. coli O157: H7 carries two copies of LPS myristoyl transferase that are differentially regulated. In this study, a whole genome-scale transcriptome specific to E. coli O157:H7 was carried out for identification of the genes differentially expressed in the amyristoylated E. coli O157:H7. The results identified a total of 110 EHEC genes that were up- or down-regulated in the amyristoylated E. coli O157:H7 strain, including genes associated with virulence (26.36%), metabolism (20.91%), transport (10.91%), signal transduction (4.55%), genetic information processing (3.64%), stress response (2.73%), regulatory function (2.73%), motility/adherence (3.64%), cell envelope (2.73%), cell division (1.82%) and ORFs of unknown function (17.27%). Of particular interest, the expression of LEE pathogenicity island genes was significantly influenced by LPS structural defects.