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Botany JoVE (Open Access) Citable · DOI

Genomic Transformation of the Picoeukaryote Ostreococcus tauri

DOI: 10.3791/4074-v
What you'll learn
  • ✓ Prepare Ostreococcus tauri cells for electroporation transformation
  • ✓ Execute electroporation protocol to introduce foreign DNA into algal cells
  • ✓ Select and identify successfully transformed colonies on selective medium
  • ✓ Troubleshoot transformation efficiency and colony appearance
Protocol

This article describes genetic transformation of the unicellular marine alga Ostreococcus tauri by electroporation. This eukaryotic organism is an effective model platform for higher plants, possesing greatly reduced genomic and cellular complexity and being readily amenable to both cell culture and chemical biology.

Difficulty
intermediate
Total time
~1–2 weeks (including culture preparation, transformation, and colony selection)
Model organism
Ostreococcus tauri
Biosafety
BSL-1

Steps

1
Prepare algal culture material for transformation

Culture and harvest Ostreococcus tauri cells in appropriate growth medium, achieving optimal cell density and physiological state for competence prior to electroporation.

▶ 01:51
2
Perform electroporation with plasmid DNA

Mix prepared algal cells with plasmid DNA and apply electrical pulses via electroporation cuvette to introduce foreign DNA into cells.

▶ 04:05
3
Plate transformed cells in semi-solid medium

Transfer electroporated cells onto selective agar plates containing semi-solid recovery medium to support cell survival and colony formation.

▶ 06:54
4
Isolate and verify transformed colonies

Monitor plates for growth of resistant colonies, identify candidate transformants, and characterize colony morphology on selective medium.

▶ 08:46
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