CONSTRUCTION OF INSERTION AND DELETION MXA MUTANTS OF METHYLOBACTERIUM-EXTORQUENS AM1 BY ELECTROPORATION

Methylobacterum extorquens AM1 is a pink-pigmented facultative methylo troph which is widely used for analyzing pathways of C-1 metabolism wi th biochemical and molecular biological techniques. To facilitate this approach, we have applied a new method to construct insertion or disr uption mutants with drug resistance genes by electroporation, By using this method, mutants were obtained in four genes present in the mxa m ethylotrophy gene cluster for which the functions were unknown, mxaR, mxaS, mxaC and mxaD. These mutants were unable to grow on methanol exc ept the mutant of mxaD, which showed reduced growth on methanol. (C) 1 998 Federation of European Microbiological Societies. Published by Els evier Science B.V. All rights reserved.