Construction of insertion and deletion mxa mutants of Methylobacterium extorquens AM1 by electroporation.
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.
Toyama
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Department of Biological Chemistry, Yamaguchi University, Japan. toyama@agr.yamaguchi-u.ac.jp
Anthony
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Lidstrom
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ME
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D002874:Chromosome Mapping; D004269:DNA, Bacterial / Q000235:genetics; D018274:Electroporation; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D016946:Gram-Negative Aerobic Rods and Cocci / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D000432:Methanol / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D016254:Mutagenesis, Insertional; D009154:Mutation; D010957:Plasmids / Q000235:genetics; D011995:Recombination, Genetic; D017384:Sequence Deletion
D004269:DNA, Bacterial; D000432:Methanol