Summary
A protocol for producing competent Pseuclomonas aeruginosa, Pseudomonas putida, and Xanthomonas maltophilia was adapted and modified from existing methods. Cells were incubated on ice for 30 minutes in buffered 100 mM MgCl2 followed by 30 minutes in buffered 100 mM CaCl2 prior to addition of DNA. The MgCl2-CaCl2 incubation increased transformation efficiency two to three times, compared with protocols which use incubation in either Mg2+ or Ca2+, but not both.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Bagdasarian, M., Lurz, R., Rückert, B., Franklin, F.C.H., Bagdasarian, M.M., Frey, J., and Timmis, K.N. (1981). Gene 16, 237–247.
Bagdasarian, M., Bagdasarian, M.M., Coleman, S., and Timmis, K.N. (1979). New vector plasmids for cloning in Pseudomonas. In: Plasmids of Medical, Environmental and Commercial Importance, K.N. Timmis and A. Pühler, eds. pp. 411–422, Amsterdam: Elsevier/North-Holland.
Dikshit, K.L. and Webster, D.A. (1988). Gene 70, 377–386.
Diver, J.M., Bryan, L.E. and Sokol, P.A. (1990). Anal. Biochem. 189, 75–79.
Liu, S.-C, Webster, D.A. and Stark, B.C. (1995). Appl. Microbiol. Biotechnol. 44, 419–424.
Mercer, A.A., and Loutit, J.S. (1979). J. Bacteriol. 140, 37–42.
Messing, J. (1983). New M13 vectors for cloning, In: Methods in Enzymology, R. Wu, L. Grossman, and K. Moldave, eds. vol. 101, pp 20–78, New York: Academic Press.
Sambrook, J., Fritsch, E.F., and Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd edn., Cold Spring Harbor, New York: Cold Spring Harbor Press.
Sano, Y. and Kageyama, M. (1977). J. Gen. Appl. Microbiol. 23, 183–186.
Sinclair, M.I. and Morgan, A.F. (1978). Aust. J. Biol. Sci. 31, 679–688.
Smith, H.O., Danner, D.B. and Deich, R.A. (1981). Ann. Rev. Biochem. 50, 41–68.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Liu, SC., Webster, D.A. & Stark, B.C. An improved method of transformation in Pseudomonads. Biotechnol Tech 10, 683–686 (1996). https://doi.org/10.1007/BF00168480
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00168480