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Identification and characterization of P GCW14 : a novel, strong constitutive promoter of Pichia pastoris

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Abstract

The available promoters in the Pichia pastoris expression platform are still limited. We selected and identified a novel strong constitutive promoter, P GCW14 , and tested its promoter activity using enhanced green fluorescent protein (EGFP) as a reporter. Potential promoter regions of P GCW14 were cloned upstream of the EGFP gene and promoter activity was analyzed by measuring fluorescence intensity. P GCW14 exhibited significantly stronger promoter activity than the classic strong constitutive promoters P TEF1 and P GAP under various carbon sources, suggesting that P GCW14 is a strong and constitutive promoter. Hence, P GCW14 can be used as a promoter for high-level expression of heterologous proteins.

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Acknowledgments

This work was supported by National Natural Science Foundation of China (No. 20976062 and 31000027), China Postdoctoral Science Foundation Grant (No. 2011M501330) and the Ministry of Science and Technology of the People’s Republic of China (National “863” Project No. 2011AA100905).

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Authors and Affiliations

  1. School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, Guangdong, China

    Shuli Liang, Chengjuan Zou, Ying Lin, Xuanwei Zhang & Yanrui Ye

  2. College of Environmental Science and Engineering, South China University of Technology, Guangzhou, 510006, Guangdong, China

    Yanrui Ye

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  1. Shuli Liang
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  2. Chengjuan Zou
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  3. Ying Lin
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  4. Xuanwei Zhang
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  5. Yanrui Ye
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Corresponding author

Correspondence to Yanrui Ye.

Additional information

Shuli Liang and Chengjuan Zou contributed equally to this study.

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Liang, S., Zou, C., Lin, Y. et al. Identification and characterization of P GCW14 : a novel, strong constitutive promoter of Pichia pastoris . Biotechnol Lett 35, 1865–1871 (2013). https://doi.org/10.1007/s10529-013-1265-8

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  • DOI: https://doi.org/10.1007/s10529-013-1265-8

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