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Applied and Environmental Microbiology, May 2002, p. 2619-2623, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2619-2623.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Cloning of Escherichia coli lacZ and lacY Genes and Their Expression in Gluconobacter oxydans and Acetobacter liquefaciens

Hesham E. Mostafa,,{dagger} Knut J. Heller,* and Arnold Geis

Institute for Microbiology, Federal Dairy Research Centre, 24103 Kiel, Germany

Received 5 December 2001/ Accepted 28 February 2002

An efficient transformation protocol for Gluconobacter oxydans and Acetobacter liquefaciens strains was developed by preparation of electrocompetent cells grown on yeast extract-ethanol medium. Plasmid pBBR122 was used as broad-host-range vector to clone the Escherichia coli lacZY genes in G. oxydans and A. liquefaciens. Although both lac genes were functionally expressed in both acetic acid bacteria, only a few transformants were able to grow on lactose. However, this ability strictly depended on the presence of a plasmid expressing both lac genes. Mutations in the plasmids and/or in the chromosome were excluded as the cause of growth ability on lactose.

* Corresponding author. Mailing address: Institute for Microbiology, Federal Dairy Research Centre, P.O. Box 6069, D-24121 Kiel, Germany. Phone: 49-(0)431-609 2340. Fax: 49-(0)431-609 2306. E-mail: heller{at}bafm.de.

{dagger} Present address: Mubarak City for Scientific Research, Institute for Genetic Engineering, Research Area-Borg El Arab, Post Code 21934, Alexandria, Egypt.

Applied and Environmental Microbiology, May 2002, p. 2619-2623, Vol. 68, No. 5
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.5.2619-2623.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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