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Applied and Environmental Microbiology, January 2005, p. 407-416, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.407-416.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Isolation and Characterization of a Native Composite Transposon, Tn14751, Carrying 17.4 Kilobases of Corynebacterium glutamicum Chromosomal DNA

Masayuki Inui,1 Yota Tsuge,1,2 Nobuaki Suzuki,1 Alain A. Vertès,1 and Hideaki Yukawa1,2*

Microbiology Research Group, Research Institute of Innovative Technology for the Earth, Soraku-Gun, Kyoto,1 Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Nara, Japan2

Received 5 July 2004/ Accepted 10 August 2004

A native composite transposon was isolated from Corynebacterium glutamicum ATCC 14751. This transposon comprises two functional copies of a corynebacterial IS31831-like insertion sequence organized as converging terminal inverted repeats. This novel 20.3-kb element, Tn14751, carries 17.4 kb of C. glutamicum chromosomal DNA containing various genes, including genes involved in purine biosynthesis but not genes related to bacterial warfare, such as genes encoding mediators of antibiotic resistance or extracellular toxins. A derivative of this element carrying a kanamycin resistance cassette, minicomposite Tn14751, transposed into the genome of C. glutamicum at an efficiency of 1.8 x 102 transformants per µg of DNA. Random insertion of the Tn14751 derivative carrying the kanamycin resistance cassette into the chromosome was verified by Southern hybridization. This work paves the way for realization of the concept of minimum genome factories in the search for metabolic engineering via genome-scale directed evolution through a combination of random and directed approaches.

* Corresponding author. Mailing address: Microbiology Research Group, Research Institute of Innovative Technology for the Earth, Soraku-Gun, Kyoto 619-0292, Japan. Phone: (81)-774-75-2308. Fax: (81)-774-75-2321. E-mail: mmg-lab{at}rite.or.jp.

Applied and Environmental Microbiology, January 2005, p. 407-416, Vol. 71, No. 1
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.1.407-416.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Vertes, A. A., Inui, M., Yukawa, H. (2005). Manipulating Corynebacteria, from Individual Genes to Chromosomes. Appl. Environ. Microbiol. 71: 7633-7642 [Full Text]  
  • Suzuki, N., Nonaka, H., Tsuge, Y., Inui, M., Yukawa, H. (2005). New Multiple-Deletion Method for the Corynebacterium glutamicum Genome, Using a Mutant lox Sequence. Appl. Environ. Microbiol. 71: 8472-8480 [Abstract] [Full Text]  
  • Suzuki, N., Okayama, S., Nonaka, H., Tsuge, Y., Inui, M., Yukawa, H. (2005). Large-Scale Engineering of the Corynebacterium glutamicum Genome. Appl. Environ. Microbiol. 71: 3369-3372 [Abstract] [Full Text]  


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