Isolation of auxotrophic mutants of Methylophilus methylotrophus by modified-marked exchange
A method of stabilizing a transposon (Tn5) has been developed which allows the isolation of stable auxotrophic mutants of Methylophilus methylotrophus AS1. Insertion of Tn5 into a cloned M. metylotrophus AS1 DNA fragment encoding anthranilate synthase followed by transfer of the vector with the modified trpE gene to M. methylotrophus AS1 resulted in unstable auxotrophs among the recombinants. Deletion of IS50R, which encodes transposase production from Tn5, stabilized the transposon after mobilization to M. methylotrophus AS1. When trpE genes with the modified Tn5 inserts were mobilized into M. methylotrophus AS1, stable, kanamycin-resistant tryptophan auxotrophs were obtained by double-crossover homologous recombination with the chromosome.
- Research Organization:
- Univ. of Minnesota, Navarre
- OSTI ID:
- 5428843
- Journal Information:
- Appl. Environ. Microbiol.; (United States), Journal Name: Appl. Environ. Microbiol.; (United States) Vol. 54:1; ISSN AEMID
- Country of Publication:
- United States
- Language:
- English
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