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Random segment deletion based on IS31831 and Cre/loxP excision system in Corynebacterium glutamicum

Abstract

A simple and random genome deletion method combining insertion sequence (IS) element IS31831 and the Cre/loxP excision system generated 42 Corynebacterium glutamicum mutants (0.2–186 kb). A total of 393.6 kb (11.9% of C. glutamicum R genome) coding for 331 genes was confirmed to be nonessential under standard laboratory conditions. The deletion strains, generated using only two vectors, varied not only in their lengths but also the location of the deletion along the C. glutamicum R genome. By comparing and analyzing the generated deletion strains, identification of nonessential genes, the roles of genes of hitherto unknown function, and gene–gene interactions can be easily and efficiently determined.

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Acknowledgment

We thank Prof. R. H. Doi (University of California at Davis) and Dr. C. Omumasaba (RITE) for critical reading of the manuscript. We are grateful to Dr. H. Nonaka (RITE, present Mie University) for providing us the useful website about C. glutamicum R genome information. This study was carried out as a part of the Project for Development of a Technological Infrastructure for Industrial Bioprocesses by the Ministry of Economy, Trade and Industry and was funded by the New Energy and Industrial Technology Development Organization.

Author information

Correspondence to Hideaki Yukawa.

Electronic supplementary material

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Supplementary Table 1

Primers used for verification of deletion (DOC 44 kb)

Supplementary Table 2

Deleted genes (DOC 100 kb)

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Tsuge, Y., Suzuki, N., Inui, M. et al. Random segment deletion based on IS31831 and Cre/loxP excision system in Corynebacterium glutamicum . Appl Microbiol Biotechnol 74, 1333–1341 (2007). https://doi.org/10.1007/s00253-006-0788-5

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Keywords

  • Corynebacterium glutamicum
  • Cre/loxP
  • IS element
  • Genomic engineering
  • Random genome deletion