Construction of Insertion and Deletion Mxa Mutants of Methylobacterium Extorquens AM1 by Electroporation

FEMS Microbiol Lett. 1998 Sep 1;166(1):1-7. doi: 10.1111/j.1574-6968.1998.tb13175.x.

Abstract

Methylobacterium extorquens AM1 is a pink-pigmented facultative methylotroph which is widely used for analyzing pathways of C1 metabolism with biochemical and molecular biological techniques. To facilitate this approach, we have applied a new method to construct insertion or disruption mutants with drug resistance genes by electroporation. By using this method, mutants were obtained in four genes present in the mxa methylotrophy gene cluster for which the functions were unknown, mxaR, mxaS, mxaC and mxaD. These mutants were unable to grow on methanol except the mutant of mxaD, which showed reduced growth on methanol.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alleles
  • Amino Acid Sequence
  • Base Sequence
  • Chromosome Mapping
  • DNA, Bacterial / genetics
  • Electroporation
  • Genes, Bacterial
  • Genetic Complementation Test
  • Gram-Negative Aerobic Rods and Cocci / genetics*
  • Gram-Negative Aerobic Rods and Cocci / growth & development
  • Gram-Negative Aerobic Rods and Cocci / metabolism
  • Methanol / metabolism
  • Molecular Sequence Data
  • Multigene Family
  • Mutagenesis, Insertional
  • Mutation*
  • Plasmids / genetics
  • Recombination, Genetic
  • Sequence Deletion

Substances

  • DNA, Bacterial
  • Methanol