Open AccessCloning of Escherichia coli lacZ and lacY Genes and Their Expression in Gluconobacter oxydans and Acetobacter liquefaciens
Author(s) -
Hesham Mostafa,
Knut J. Heller,
Arnold Geis
Publication year - 2002
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.68.5.2619-2623.2002
Subject(s) - plasmid , biology , escherichia coli , transformation (genetics) , acetobacter , cloning (programming) , acetic acid bacteria , gene , lac operon , microbiology and biotechnology , bacteria , clone (java method) , lactose , expression vector , genetics , biochemistry , recombinant dna , computer science , programming language
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.