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Novel synthetic signal peptides for the periplasmic secretion of green fluorescent protein in Escherichia coli

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Abstract

New secretion vectors containing synthetic signal peptides were constructed to study the periplasmic translocation of green fluorescent protein (GFP) in Escherichia coli. These constructs encode synthetic signal peptides spA and spD fused to the amino terminal end of GFP, and expressed from T7/lac promoter in the BL21DE3 strain by induction with IPTG. The recombinant protein was detected in both the cytoplasmic and periplasmic fractions. Fluorescence analysis revealed that recombinant proteins with signal peptides were not fluorescent, indicating translocation to periplasmic space. In contrast, recombinant proteins without signal peptide were fluorescent. These results indicate that the expressed recombinant proteins were translocated into the periplasm. Therefore, the synthetic signal peptides derived from signal peptides of Bacillus sp. could efficiently secrete the heterologous proteins to the periplasmic space of E. coli.

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Acknowledgment

We would like to thank the Malaysian Genome Institute (MGI), Ministry of Science, Technology and Innovation, for funding this study under the Genomics and Molecular Biology Initiatives Programme (Project num 07-05-MGI-GMB011).

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Vithya Velaithan & Raha Abdul Rahim

  2. Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Sieo Chin Chin & Khatijah Yusoff

  3. Institute of Bioscience, Universiti Putra Malaysia, 43400, Serdang, Selangor, Malaysia

    Sieo Chin Chin, Khatijah Yusoff & Raha Abdul Rahim

  4. Department of Bioprocess Engineering, Faculty of Chemical and Natural Resources Engineering, Universiti Teknologi Malaysia, Skudai, Johor, Malaysia

    Rosli Md. Illias

Authors
  1. Vithya Velaithan
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  2. Sieo Chin Chin
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  3. Khatijah Yusoff
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  4. Rosli Md. Illias
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  5. Raha Abdul Rahim
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Corresponding author

Correspondence to Raha Abdul Rahim.

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Velaithan, V., Chin, S.C., Yusoff, K. et al. Novel synthetic signal peptides for the periplasmic secretion of green fluorescent protein in Escherichia coli . Ann Microbiol 64, 543–550 (2014). https://doi.org/10.1007/s13213-013-0687-9

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  • DOI: https://doi.org/10.1007/s13213-013-0687-9

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