Secretion of biologically active human epidermal growth factor from Escherichia coli using Yersinia pestis Caf1 signal peptide

Yi Lin Liu, Liang Min Huang, Wen Po Lin, Chung Chin Tsai, Tsung Shun Lin, Yu Hua Hu, Hquan Shin Chen, Jun Ming Han, Hsian-Jenn Wang, Yu Tien Liu

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Background and Purpose: The Caf1 secretion pathway of Yersinia pestis is one of the most well-characterized export machineries. To facilitate the secretion of human epidermal growth factor (hEGF) in Escherichia coli, a DNA fragment containing the synthetic gene for hEGF was joined to a sequence encoding the signal peptide of Yersinia pestis Caf1 protein. Methods: The gene for hEGF was synthesized by overlapping polymerase chain reaction technique and was placed under the control of the caf1 gene promoter in the recombinant plasmid pHL401 which was used to transfect E. coli BL-21 for production of hEGF. The biological function of recombinant hEGF was measured by estimating its ability to stimulate the proliferation of human embryonic kidney-293 cells. Results: The results indicated that the expressed hybrid protein was processed during the secretion process. The majority of the mature hEGF was recovered from the periplasm and medium fractions, with a small amount of the expressed hEGF deposited in the cytoplasm. Furthermore, it was found that the cell proliferation was enhanced by the recombinant hEGF. Conclusion: These results suggested that the recombinant hEGF was successfully secreted through the inner membrane of cells into the periplasm and then through the outer membrane into the medium via the action of the signal peptide of Y. pestis Caf1 in E. coli. The mitogenic activity of hEGF in cells was demonstrated.

Original languageEnglish
Pages (from-to)366-371
Number of pages6
JournalJournal of Microbiology, Immunology and Infection
Volume39
Issue number5
Publication statusPublished - Oct 1 2006
Externally publishedYes

Keywords

  • Bacterial proteins
  • Caf1
  • Protein sorting signals
  • Recombinant fusion proteins
  • Yersinia pestis

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology and Microbiology(all)
  • Microbiology (medical)
  • Infectious Diseases

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  • Cite this

    Liu, Y. L., Huang, L. M., Lin, W. P., Tsai, C. C., Lin, T. S., Hu, Y. H., Chen, H. S., Han, J. M., Wang, H-J., & Liu, Y. T. (2006). Secretion of biologically active human epidermal growth factor from Escherichia coli using Yersinia pestis Caf1 signal peptide. Journal of Microbiology, Immunology and Infection, 39(5), 366-371.