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Chromosomal integration of the Vitreoscilla hemoglobin gene and its physiological actions in Tremella fuciformis

Abstract

The Vitreoscilla hemoglobin (VHb) gene was expressed in yeast-like conidia (YLCs) of Tremella fuciformis (T. fuciformis) to increase cell density in submerged fermentation by enhancing oxygen uptake. With the intention of doing this, an integrated expression vector containing the VHb gene and the hygromycin B phosphotransferase (hph) gene derived from Escherichia coli (E. coli) as the selectable marker was constructed, and then transformed into protoplasts of YLCs from T. fuciformis with restriction enzyme-mediated DNA integration (REMI). Hygromycin-resistant transformants had been generated during the transformation. Molecular evidences including PCR assay, Southern blotting, and Western blot analysis indicated the VHb gene had been integrated into the genome of transgenic T. fuciformis strains and was expressed successfully. Shake-flask fermentation and bioreactor cultivation results showed that the expression of VHb in this fungus could enhance growth of YLCs. The final cell density was higher in the culture of VHb-expressing strain than that of the wild-type strain. Moreover, these results also suggested that CaMV35S promoter was capable of driving the expression of heterologous genes in T. fuciformis.

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Acknowledgements

We wish to thank Dr. Xu ZP of Zhejiang University for the kindness to supply all bacterial plasmids. We also thank sincerely MD Chen LC for critical reading of the manuscript and useful suggestions.

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Correspondence to Ya-Ling Shen or Dong-Zhi Wei.

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Zhu, H., Wang, T., Sun, S. et al. Chromosomal integration of the Vitreoscilla hemoglobin gene and its physiological actions in Tremella fuciformis . Appl Microbiol Biotechnol 72, 770–776 (2006). https://doi.org/10.1007/s00253-006-0322-9

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Keywords

  • Fermentation
  • Southern Hybridization
  • Submerged Fermentation
  • Final Cell Density
  • Increase Cell Density