TY - JOUR AU - Hobson, Neil AU - Price, Nancy L. AU - Ward, Jordan D. AU - Raivio, Tracy L. PY - 2008 DA - 2008/08/05 TI - Generation of a restriction minus enteropathogenic Escherichia coli E2348/69 strain that is efficiently transformed with large, low copy plasmids JO - BMC Microbiology SP - 134 VL - 8 IS - 1 AB - Many microbes possess restriction-modification systems that protect them from parasitic DNA molecules. Unfortunately, the presence of a restriction-modification system in a given microbe also hampers genetic analysis. Although plasmids can be successfully conjugated into the enteropathogenic Escherichia coli strain E2348/69 and optimized protocols for competent cell preparation have been developed, we found that a large, low copy (~15) bioluminescent reporter plasmid, pJW15, that we modified for use in EPEC, was exceedingly difficult to transform into E2348/69. We reasoned that a restriction-modification system could be responsible for the low transformation efficiency of E2348/69 and sought to identify and inactivate the responsible gene(s), with the goal of creating an easily transformable strain of EPEC that could complement existing protocols for genetic manipulation of this important pathogen. SN - 1471-2180 UR - https://doi.org/10.1186/1471-2180-8-134 DO - 10.1186/1471-2180-8-134 ID - Hobson2008 ER -