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Science 11 October 1985:
Vol. 230. no. 4722, pp. 179 - 181
DOI: 10.1126/science.2994228
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Articles

Science, Vol 230, Issue 4722, 179-181
Copyright © 1985 by American Association for the Advancement of Science

articles

Cloning of Shiga-like toxin structural genes from a toxin converting phage of Escherichia coli

JW Newland, NA Strockbine, SF Miller, AD O'Brien, and RK Holmes

The genes controlling high-level production of Shiga-like toxin (SLT) in Escherichia coli were cloned from the SLT converting phage 933J. This phage was isolated from a strain of E. coli that caused a foodborne outbreak of hemorrhagic colitis. The genes that convert normal E. coli to organisms producing high levels of toxin were cloned into the plasmid pBR328 and expressed in E. coli HB101. DNA restriction mapping, subcloning, examination of the cloned gene products by minicell analysis, neutralization, and immunoprecipitation with antibodies to SLT were used to localize the toxin converting genes and identify them as structural genes for SLT. Southern hybridization studies established that the DNA fragment carrying the cloned toxin structural genes had homology with the DNA of Shigella.


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