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A novel expression vector for the cyanobacterium, Synechococcus PCC 6301

Y Takeshima1, M Sugiura, H Hagiwara

  • 1Hagiwara Institute of Health, Japan.

DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes
|January 1, 1994
PubMed
Summary

Researchers developed a new cyanobacterial expression vector, pARUB19, for stable foreign gene expression. This vector enables luciferase gene insertion and expression in Synechococcus PCC 6301, facilitating biotechnology applications.

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Area of Science:

  • Molecular Biology
  • Biotechnology
  • Microbiology

Background:

  • Cyanobacteria are photosynthetic microorganisms with potential for biotechnological applications.
  • Efficient expression systems are crucial for harnessing the capabilities of cyanobacteria.
  • Developing stable expression vectors is key for genetic manipulation of cyanobacteria.

Purpose of the Study:

  • To construct and characterize a novel cyanobacterial expression vector, pARUB19.
  • To demonstrate the stable maintenance and expression of foreign genes in Synechococcus PCC 6301.
  • To assess the potential of this vector for high-level foreign gene expression.

Main Methods:

  • Construction of a recombinant plasmid (pARUB19) using RuBisCO promoter/terminator sequences from Synechococcus PCC 6301.

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  • Inclusion of an ampicillin-resistance gene (ApR) as a selectable marker and unique restriction sites for gene insertion.
  • Transformation of Synechococcus PCC 6301 with the luciferase gene from Photinus pyralis using the constructed vector.

    • Main Results:

    • The pARUB19 vector was successfully constructed and maintained stably in Synechococcus PCC 6301 host cells.
    • Expression of the firefly luciferase gene was confirmed by light production in transformant cells.
    • Luciferase constituted 1.2% of the total soluble protein in the transformed cyanobacteria.

    Conclusions:

    • The developed cyanobacterial expression vector, pARUB19, is effective for stable foreign gene expression in Synechococcus PCC 6301.
    • This system shows promise for facilitating higher levels of foreign protein production in cyanobacteria.
    • The pARUB19 vector represents a valuable tool for genetic engineering and synthetic biology in cyanobacteria.