Biotechnology Letters

, Volume 26, Issue 21, pp 1659–1663

Construction of a pTOC-T vector using GST-ParE toxin for direct cloning and selection of PCR products

  • Han Geun Kim
  • Hye Sun Kim
  • Hyun Jin Hwang
  • Sung Kyun Chung
  • Jung Min Lee
  • Dae Kyun Chung
Article

DOI: 10.1007/s10529-004-3518-z

Cite this article as:
Kim, H.G., Kim, H.S., Hwang, H.J. et al. Biotechnol Lett (2004) 26: 1659. doi:10.1007/s10529-004-3518-z

Abstract

Using linker insertion mutations, we determined the most stable region of the parE gene which encodes a toxic protein (ParE) that inhibits growth of Escherichia coli. The toxicity of ParE was sustained until a 144 bp linker was inserted into this region. We have developed a 3′ T-overhang vector based on these characteristics of the GST-ParE toxin, and named pTOC-T. Because pTOC-T uses a post-segregational killing system, all transformants grown up on the plates can be considered as recombinants containing foreign DNA. pTOC-T not require X-Gal, IPTG or other substrates for selection. This T-vector using a positive selection system can be applied to various E. coli strains such as XL1-Blue, BL21, DH5α, JM109, and JM110.

Keywords

bacterial toxin broad host range cloning vector ParE protein positive selection vector T-vector 

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Han Geun Kim
    • 1
  • Hye Sun Kim
    • 1
  • Hyun Jin Hwang
    • 2
  • Sung Kyun Chung
    • 2
  • Jung Min Lee
    • 1
  • Dae Kyun Chung
    • 1
    • 2
  1. 1.School of Biotechnology and Institute of Life Science and ResourcesKyung Hee UniversitySuwon,Korea
  2. 2.RNA Inc.Suwon,Korea

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