Use of transmissible plasmids as cloning vectors in Caulobacter crescentus

Gene
P V SchoenleinB Ely

Abstract

Cloning vectors for studies of Caulobacter crescentus genes should be transferrable between Escherichia coli and C. crescentus since a transformation system has not been developed for C. crescentus. We have tested a large number of vectors containing IncP or IncQ replicons and found that many of the vectors containing IncQ replicons, and all but one of the vectors containing IncP replicons, are readily transferred by conjugation into C. crescentus. All of the plasmids tested were maintained in C. crescentus at 1 to 5 copies per cell, but plasmids containing IncP replicons were more stable than plasmids containing IncQ replicons. Further studies with a derivative of the IncQ plasmid R300B showed that when a promoterless kanamycin (Km)-resistance gene (npt2) was inserted into the intercistronic region of the sul-aphC (SuR-SmR) operon, Km resistance was expressed only when the npt2 gene was inserted such that it would be transcribed from the sul promoter. These data indicate that R300B does not contain sequences which would provide promoter function in C. crescentus in the orientation opposite to that of the sul operon and that any genes cloned in this orientation would require native promoters for expression. To provide greater v...Continue Reading

References

Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D H Figurski, D R Helinski
Jun 1, 1976·Journal of Bacteriology·N B WoodL Shapiro
Jan 1, 1985·Molecular & General Genetics : MGG·B Ely
Mar 5, 1987·Journal of Molecular Biology·K M Hahnenberger, L Shapiro
Oct 1, 1984·Journal of Bacteriology·M E WinklerB Ely
Mar 1, 1981·Journal of Bacteriology·D O WoodM B Tindol
Feb 1, 1982·Journal of Bacteriology·B Ely, R H Croft
Dec 5, 1981·Journal of Molecular Biology·N Ohta, A Newton
Aug 1, 1982·Journal of Bacteriology·J T BarrettB Ely
Nov 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·M MilhausenN Agabian
Aug 1, 1983·Analytical Biochemistry·J SummertonR Bestwick
Mar 1, 1982·Journal of Bacteriology·J T BarrettB Ely
Dec 1, 1980·Proceedings of the National Academy of Sciences of the United States of America·G DittaD R Helinski
Nov 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·M PuruckerL Shapiro
Nov 1, 1977·Genetics·B Ely, R C Johnson

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Citations

Sep 5, 2001·American Journal of Human Biology : the Official Journal of the Human Biology Council·A. Fernández-SantanderP. Moral
Jan 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·S P WangB Ely
Sep 28, 1998·Journal of Bacteriology·G LeclercB Ely

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