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Construction of genetic parts from the Corynebacterium glutamicum genome with high expression activities

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Abstract

Objective

To construct effective genetic expression parts controlling transcription and translation initiation for synthetic biology and heterologous expression in Corynebacterium glutamicum.

Result

Twelve highly expressed genes were identified from the proteomic data of C. glutamicum. Their related sequences were used to construct bicistronic genetic expression parts. Each part contain promoter, 5′-UTR, N-terminal sequence of the source gene and a conserved SD sequence, associated with target gene, forming the bicistronic expression cassette. The enhanced green fluorescent protein (EGFP) expression levels controlled by these novel parts have 1.4 to 790-fold increase in C. glutamicum compared with corresponding promoter-5′-UTR part. One of the bicistronic parts is 1.35 times the EGFP expression of the constitutive-expression pXMJ19. These bicistronic parts had expression advantage compared with conventional promoter-5′-UTR parts.

Conclusion

Various genetic parts for efficient gene expression can be quickly obtained via this new method.

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Acknowledgment

This work was financially supported by the National Basic Research Program of China (973 Program) (Grant No. 2013CB733602), the Fundamental Research Funds for the Central Universities (Grant No. JUSRP51401A) and the Natural Science Foundation of Jiangsu Province (Grant No. BK20150148).

Supporting information

Supplementary Table 1—Primers used.

Supplementary Table 2—Information about highly expressed genes in C. glutamicum.

Supplementary Fig. 1—Construction process of the probe vector pE-0.

Supplementary Fig. 2—Insertion of fragments guided by Golden Gate method to seamlessly connect genetic expression parts to reporter gene.

Author information

Authors and Affiliations

  1. National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi, 214122, China

    Zihao Zhao, Xiuxia Liu, Wei Zhang, Yankun Yang, Xiaofeng Dai & Zhonghu Bai

  2. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China

    Zihao Zhao, Xiuxia Liu, Wei Zhang, Yankun Yang, Xiaofeng Dai & Zhonghu Bai

Authors
  1. Zihao Zhao
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  2. Xiuxia Liu
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  3. Wei Zhang
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  4. Yankun Yang
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  5. Xiaofeng Dai
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  6. Zhonghu Bai
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Corresponding authors

Correspondence to Xiuxia Liu or Zhonghu Bai.

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The authors declare that they have no conflict of interest.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Zhao, Z., Liu, X., Zhang, W. et al. Construction of genetic parts from the Corynebacterium glutamicum genome with high expression activities. Biotechnol Lett 38, 2119–2126 (2016). https://doi.org/10.1007/s10529-016-2196-y

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  • DOI: https://doi.org/10.1007/s10529-016-2196-y

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