Isolation of Auxotrophic Mutants of Methylophilus methylotrophus by Modified-Marker Exchange-Reference-Cited by-同舟云学术

Isolation of Auxotrophic Mutants of Methylophilus methylotrophus by Modified-Marker Exchange

Published:1988-01 Issue:1 Volume:54 Page:271-273
ISSN:0099-2240
Container-title:Applied and Environmental Microbiology
language:en
Short-container-title:Appl Environ Microbiol

Author:

Bohanon Michael J.¹,Bastien Cathryn A.¹,Yoshida Roberta¹,Hanson Richard S.¹

Affiliation:

1. Gray Freshwater Biological Institute, University of Minnesota, Navarre, Minnesota 55392

Abstract

A method for stabilizing a transposon (Tn 5 ) has been developed which allows the isolation of stable auxotrophic mutants of Methylophilus methylotrophus ASI. Insertion of Tn 5 into a cloned M. methylotrophus ASI DNA fragment encoding anthranilate synthase followed by transfer of the vector with the modified trpE gene to M. methylotrophus ASI resulted in unstable auxotrophs among the recombinants. Deletion of IS 50 R, which encodes transposase production from Tn 5 , stabilized the transposon after mobilization to M. methylotrophus ASI. When trpE genes with the modified Tn 5 inserts were mobilized into M. methylotrophus ASI, stable, kanamycin-resistant tryptophan auxotrophs were obtained by double-crossover homologous recombination with the chromosome.

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

Link

https://journals.asm.org/doi/pdf/10.1128/aem.54.1.271-273.1988

Reference10 articles.

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