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KMID : 0380420010250030133
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Journal of Prventive Veterinary Medicine
2001 Volume.25 No. 3 p.133 ~ p.140
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Expression and Characteristics of C-terminal Portions of Escherichia coli O157:H7 Intimin
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Gannon, V. P. J.
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Abstract
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Portions of the intimin genes of Escherichia coli O157:H7 strain 319 and of the enteropathogenic E. coli O127:H6 strain E2348/69 were amplified by PCR and cloned into pET-28a (+) expression vectors. The entire 934 aa of E. coli O157:H7 intimin, the C-terminal 306 aa of E. coli O157:H7 intimin, and the C-terminal 311 aa of E. coli O127:H6 intimin were expressed as protein fusions with a six histidine residue tag (His-tag) in pET-28a (+). Rabbit antisera raised against the six His-tag 3´ end one-third portion of E. coli O157:H7 intimin protein fusion reacted strongly in Western blots with original antigen and its homology protein, the six His-tag-fusioned protein containing full-length intimin of E. coli O157:H7, while reacting weakly with the his-fusioned intimin from enteropathogenic E. coli serotype O127:H6. In contrast with the antiseraraised against the sis His-tag 3´end portion of E. coli O127:H6 recognized only the original antigen, His-intO127C.
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