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Stable RK2-Derived Cloning Vectors for the Analysis of Gene Expression and Gene Function in Gram-Negative Bacteria

March 2001 , Volume 14 , Number  3
Pages  426 - 430

Bruno Dombrecht , Jos Vanderleyden , and Jan Michiels

F.A. Janssens Laboratory of Genetics, Katholieke Universiteit Leuven, B-3001 Heverlee, Belgium

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Accepted 29 November 2000.

The construction of several stable RK2-derived cloning vectors for the analysis of gene expression and function in gram-negative bacteria is reported. Plasmid stability is conferred by the RK2 par locus or by insertion of the spsAB or spsCD symbiotic plasmid stability loci from pNGR234a of Rhizobium sp. NGR234. The vectors carry multiple cloning sites with protection against read-through transcriptional activity of vector sequences. Vector derivatives with the constitutive nptII promoter or a promoter-less gusA gene are suitable for the study of gene function or regulation in bacteria.

Additional keywords: Azospirillum brasilense Sp245, post-segregational killing, pTR102, Rhizobium etli CNPAF512, y4dLM, y4mFE.

© 2001 The American Phytopathological Society