Construction and characterization of mutant Enteroaggregative E. coli O42 strain with high-pathogenicity island deletion

Abstract

Abstract: Objective To construct and characterize a mutant Enteroaggregative E. coli(EAggEC) O42 strain with in-frame deletion of high-pathogenicity island (HPI). Methods The kanamycin resistance gene (kan) was inserted between the sequences of irp8 and irp5 genes as the two homologous sequences for construction of the recombinant plasmid pCO85 by subcloning the recombined sequence into the suicide vector pCVD442. By homologous recombination and conjunction mobilization, EAO85 mutant with deletion of the core region of HPI about 24 kb spanning from irp8 to irp5 sequences was screened. Results Irp8 and irp5 genes of EAggEC O42 were exchanged by pCO85 by conjunction mobilization with the selection by sucrose. The EAO85 mutant was identified by their failure to yield PCR products with primers specific for the internal regions of HPI. Conclusion We have successfully constructed a mutant of EAggEC O42 strain with HPI in-frame deletion, which may facilitate the exploration of the role of HPI in EAggEC strain.

CLC Number:

R-33

HU Jing¹, YU Shou-yi¹, KAN Biao², LIU Zhi-hua³. Construction and characterization of mutant Enteroaggregative E. coli O42 strain with high-pathogenicity island deletion[J]. Journal of Southern Medical University, 2005, 25(11): 1352-1356.

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