Development of a markerless gene replacement system in Corynebacterium glutamicum using upp as a counter-selection marker

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• 作者：Wenwen Ma (1) (2) (3) (4)
  Xiaoyue Wang (1) (2) (3) (4)
  Yufeng Mao (1) (2) (3) (4)
  Zhiwen Wang (1) (2) (3) (4)
  Tao Chen (1) (2) (3) (4)
  Xueming Zhao (1) (2) (3) (4)
  
  1. Department of Biochemical Engineering ; School of Chemical Engineering and Technology ; Tianjin University ; Tianjin ; 300072 ; China
  2. Key Laboratory of Systems Bioengineering (Ministry of Education) ; Tianjin University ; Tianjin ; 300072 ; China
  3. SynBio Research Platform ; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) ; School of Chemical Engineering and Technology ; Tianjin University ; Tianjin ; 300072 ; China
  4. Edinburgh-Tianjin Joint Research Centre for Systems Biology and Synthetic Biology ; Tianjin University ; Tianjin ; 300072 ; China
  
• 关键词：Corynebacterium glutamicum ; Double ; strand break ; Markerless gene replacement ; Recombination ; I ; SceI ; upp
• 刊名：Biotechnology Letters
• 出版年：2015
• 出版时间：March 2015
• 年：2015
• 卷：37
• 期：3
• 页码：609-617
• 全文大小：504 KB
• 参考文献：1. Becker, J, Wittmann, C (2012) Bio-based production of chemicals, materials and fuels鈥擟orynebacterium glutamicum as versatile cell factory. Curr Opin Biotechnol 23: pp. 631-640 p://dx.doi.org/10.1016/j.copbio.2011.11.012" target="_blank" title="It opens in new window">CrossRef
  2. Binder, S, Siedler, S, Marienhagen, J, Bott, M, Eggeling, L (2013) Recombineering in Corynebacterium glutamicum combined with optical nanosensors: a general strategy for fast producer strain generation. Nucleic Acid Res 41: pp. 6360-6369 p://dx.doi.org/10.1093/nar/gkt312" target="_blank" title="It opens in new window">CrossRef
  3. Eggeling, L, Bott, M (2010) Handbook of Corynebacterium glutamicum. CRC Press, Boca Raton, Florida
  4. Hall, SD, Kolodner, RD (1994) Homologous pairing and strand exchange promoted by the Escherichia coli RecT protein. Proc Natl Acad Sci USA 91: pp. 3205-3209 p://dx.doi.org/10.1073/pnas.91.8.3205" target="_blank" title="It opens in new window">CrossRef
  5. Inui, M, Kawaguchi, H, Murakami, S, Vert猫s, AA, Yukawa, H (2004) Metabolic engineering of Corynebacterium glutamicum for fuel ethanol production under oxygen-deprivation conditions. J Mol Microbiol Biotechnol 8: pp. 243-254 p://dx.doi.org/10.1159/000086705" target="_blank" title="It opens in new window">CrossRef
  6. Jager, W, Schafer, A, Puhler, A, Labes, G, Wohlleben, W (1992) Expression of the Bacillus subtilis sacB gene leads to sucrose sensitivity in the gram-positive bacterium Corynebacterium glutamicum but not in Streptomyces lividans. J Bacteriol 174: pp. 5462-5465
  7. Kim, IK, Jeong, WK, Lim, SH, Hwang, IK, Kim, YH (2011) The small ribosomal protein S12P gene rpsL as an efficient positive selection marker in allelic exchange mutation systems for Corynebacterium glutamicum. J Microbiol Methods 84: pp. 128-130 p://dx.doi.org/10.1016/j.mimet.2010.10.007" target="_blank" title="It opens in new window">CrossRef
  8. Mitsuhashi, S (2014) Current topics in the biotechnological production of essential amino acids, functional amino acids, and dipeptides. Curr Opin Biotechnol 26: pp. 38-44 p://dx.doi.org/10.1016/j.copbio.2013.08.020" target="_blank" title="It opens in new window">CrossRef
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  10. Sander, JD, Joung, JK (2014) CRISPR-Cas systems for editing, regulating and targeting genomes. Nat Biotechnol 32: pp. 347-355 p://dx.doi.org/10.1038/nbt.2842" target="_blank" title="It opens in new window">CrossRef
  11. Shi, T, Wang, G, Wang, Z, Fu, J, Chen, T, Zhao, X (2013) Establishment of a markerless mutation delivery system in Bacillus subtilis stimulated by a double-strand break in the chromosome. PLoS One 8: pp. e81370 p://dx.doi.org/10.1371/journal.pone.0081370" target="_blank" title="It opens in new window">CrossRef
  12. Suzuki, N, Nonaka, H, Tsuge, Y, Okayama, S, Inui, M, Yukawa, H (2005) Multiple large segment deletion method for Corynebacterium glutamicum. Appl Microbiol Biotechnol 69: pp. 151-161 p://dx.doi.org/10.1007/s00253-005-1976-4" target="_blank" title="It opens in new window">CrossRef
  13. Suzuki, N, Tsuge, Y, Inui, M, Yukawa, H (2005) Cre/loxP-mediated deletion system for large genome rearrangements in Corynebacterium glutamicum. Appl Environ Microbiol 67: pp. 225-233
  14. Tan, Y, Xu, D, Li, Y, Wang, X (2012) Construction of a novel sacB-based system for marker-free gene deletion in Corynebacterium glutamicum. Plasmid 67: pp. 44-52 p://dx.doi.org/10.1016/j.plasmid.2011.11.001" target="_blank" title="It opens in new window">CrossRef
  15. Zhang, Y, Shang, X, Lai, S, Zhang, G, Liang, Y, Wen, T (2012) Development and application of an arabinose-inducible expression system by facilitating inducer uptake in Corynebacterium glutamicum. Appl Environ Microbiol 78: pp. 5831-5838 p://dx.doi.org/10.1128/AEM.01147-12" target="_blank" title="It opens in new window">CrossRef
  16. Zhu, N, Xia, H, Wang, Z, Zhao, X, Chen, T (2013) Engineering of acetate recycling and citrate synthase to improve aerobic succinate production in Corynebacterium glutamicum. PLoS One 8: pp. e60659 p://dx.doi.org/10.1371/journal.pone.0060659" target="_blank" title="It opens in new window">CrossRef
  17. Zhu, N, Xia, H, Yang, J, Zhao, X, Chen, T (2014) Improved succinate production in Corynebacterium glutamicum by engineering glyoxylate pathway and succinate export system. Biotechnol Lett 36: pp. 553-560 p://dx.doi.org/10.1007/s10529-013-1376-2" target="_blank" title="It opens in new window">CrossRef
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

Corynebacterium glutamicum is well-established for industrial and biotechnological applications. However, its genetic manipulation has generally lagged behind traditional genetic models. In this study, a counter-selectable marker gene upp was firstly confirmed to be more efficient than traditional sacB. Furthermore, a markerless gene replacement system was developed by combining upp with double-strand break repair caused by the exogenous endonuclease I-SceI. Finally, genetic modification using a dsDNA PCR fragment was carried out with the expression of recombinase/exonuclease RecE/RecT. Our results show that the genetic modification system allows precise and markerless gene replacement without altering the chromosome, with a simplified screening procedure to generate its modification.