RSF1010-based shuttle vectors for cloning and expression in Pasteurella multocida.

M D Lee; A D Henk

doi:10.1016/s0378-1135(96)01294-1

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RSF1010-based shuttle vectors for cloning and expression in Pasteurella multocida.

M D Lee·A D Henk

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Abstract

The broad host-range cloning vectors, pJRD215 and pMMB67EH, were evaluated for stability and cloning efficiency in Pasteurella multocida. Transformation of P. multocida by electroporation was unreliable and poorly efficient regardless of whether the transforming DNA was isolated from E. coli or P. multocida. Both vectors contain a mob site that enabled transfer by conjugation from E. coli to P. multocida with high efficiency. Kanamycin, streptomycin, and ampicillin resistance encoded by the vectors were expressed in P. multocida. LacZ was cloned in pMMB67EH, an expression vector, and was transferred to P. multocida by conjugation. The transconjugants expressed a functional beta-galactosidase as determined by o-nitrophenyl-beta-D-galactopyranoside (ONPG) test. We propose the use of these cosmid and expression vectors as a shuttle vectors for cloning in P. multocida.

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Ampicillin Resistance Anti-Bacterial Agents Cloning, Molecular Conjugation, Genetic Drug Resistance, Microbial Electroporation Escherichia coli Genetic Vectors Kanamycin Resistance Microbial Sensitivity Tests Pasteurella multocida Plasmids Recombinant Fusion Proteins Transformation, Bacterial beta-Galactosidase Journal Article

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M D Lee, A D Henk. 1997. RSF1010-based shuttle vectors for cloning and expression in Pasteurella multocida.. https://doi.org/10.1016/s0378-1135(96)01294-1

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