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Both the AOX1 promoter and the FLD1 promoter work together in a Pichia pastoris expression vector

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Abstract

We constructed a Pichia pastoris expression vector with two strongly inducible promoters (an alcohol oxidase 1 promoter and a formaldehyde dehydrogenase 1 promoter) based on pPIC9 k. To test the function of these promoters, the vector was used to co-express two genes that encode for green fluorescent protein (GFP) and a portion of a gelatin gene (an intra- and extracellular protein). The gelatin gene was placed under the control of PAOX1, while the GFP was under the control of PFLD1. The two proteins were simultaneously expressed upon induction with 0.5% (v/v) methanol. The two promoters functioned effectively and their coexistence on one vector did not affect their efficiency in protein expression. Thus, it was possible to simultaneously induce the expression of at least two proteins from one vector, using two different promoters.

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Acknowledgments

We thank Professor Tai Wang of the Institute of Botany, Chinese Academy of Sciences for his kind donation of vector pCAMBIA1302 containing the GFP gene, and Zhuyun Deng, the Institute of Botany, Chinese Academy of Sciences for her assistance in fluorescence microscopy. We also thank Haiyan Liu of Beijing University of Chemical Technology for proofreading the manuscript.

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Authors and Affiliations

  1. College of Life Science and Technology, Beijing University of Chemical Technology, 100029, Beijing, People’s Republic of China

    Huiming Duan, Sirajo Umar, Yiping Hu & Jinchun Chen

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  1. Huiming Duan
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  2. Sirajo Umar
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  3. Yiping Hu
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  4. Jinchun Chen
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Correspondence to Jinchun Chen.

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Duan, H., Umar, S., Hu, Y. et al. Both the AOX1 promoter and the FLD1 promoter work together in a Pichia pastoris expression vector. World J Microbiol Biotechnol 25, 1779–1783 (2009). https://doi.org/10.1007/s11274-009-0077-4

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  • DOI: https://doi.org/10.1007/s11274-009-0077-4

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