Use of a plasmid of a Yersinia enterocolitica biogroup 1A strain for the construction of cloning vectors

A plasmid with a size of 2682 base pairs isolated from the Yersinia enterocolitica biogroup 1A strain # 29807 was characterized in respect to its suitability as a basic replicon for cloning vectors. The copy number of the plasmid was determined to be approximately 14 copies per cell and it was shown to be compatible with vectors with an origin of replication derived from ColE1 and p15A. The replication region of the plasmid encodes a primer RNAI and countertranscript RNAII. Two vectors, pIV1 and pIV2, containing a kanamycin resistance gene and the lacZ alpha fragment with the multiple cloning site of pBluescriptSK + were constructed. A mobilizable derivative was successfully introduced into different bacteria belonging to the family Enterobacteriacea. To prove the applicability of the novel vectors for cloning purposes, a 13 kb hemolysin operon of Escherichia coli was inserted into pIV1, and the resulting recombinant plasmid was stably maintained and expressed in E. coli and Y.enterocolitica. (C) 2000 Elsevier Science B.V. All rights reserved.