Transformation of algal cells

• US 8,759,615 B2
• Filed: 02/13/2012
• Issued: 06/24/2014
• Est. Priority Date: 06/06/2008
• Status: Expired due to Fees

• Current Assignee: Aurora Algae, Incorporated (Reliance Industries Limited)
• Original Assignee: Aurora Algae, Incorporated (Reliance Industries Limited)
• Inventors: Vick, Bertrand, Kilian, Oliver
• Primary Examiner(s): Fox, David T
• Application Number:
  US13/372,467
• Publication Number:
  US 20120208279A1
• Time in Patent Office: 862 Days
• Field of Search: None
• US Class Current: 800/292
• CPC Class Codes: C12N 1/12 Unicellular algae; Culture media therefor as new plants A01H13/00
• CPC Class Codes: C12N 15/79 Vectors or expression systems specially adapted for eukaryotic hosts
• CPC Class Codes: C12N 15/8206 by physical or chemical, i.e. non-biological, means, e.g. electroporation, PEG mediated
• CPC Class Codes: C12N 15/8207 by mechanical means, e.g. microinjection, particle bombardment, silicon whiskers
• CPC Class Codes: C12N 15/8237 Externally regulated expression systems
• CPC Class Codes: C12N 15/8247 involving modified lipid metabolism, e.g. seed oil composition
• CPC Class Codes: C12N 15/90 Stable introduction of foreign DNA into chromosome

Abstract

Exemplary methods include a method for transforming an algal cell by preparing a transformation construct, preparing a particle for bombarding the algal cell, adhering the transformation construct to the particle, bombarding the algal cell with the particle, and growing the algal cell into a colony. The transformation construct is replicated within a nuclear genome of the algal cell and the growing of the algal cell is in a nutrient medium. Another exemplary method may include a method for genetically modifying an algal cell, by adding nucleic acid to the algal cell while the algal cell is suspended in a solution of low conductivity, introducing the nucleic acid into the algal cell by application of an electrical pulse resulting in a transformed algal cell, and selecting a colony that includes the transformed algal cell.

Claims

1. A method for genetically modifying an algal cell, the method comprising:

adding nucleic acid to the algal cell while the algal cell is suspended in a solution of low conductivity, the nucleic acid including a gene involved in lipid biosynthesis;

introducing the nucleic acid into the algal cell by application of an electrical pulse resulting in a transformed algal cell; and

selecting a colony that includes the transformed algal cell;

wherein the electrical pulse is applied with a field strength of higher than approximately 7500 V/cm; and

wherein the algal cell is of genus Nannochloropsis.

2. The method of claim 1, wherein the nucleic acid includes a transformation construct.

3. The method of claim 1, wherein the electrical pulse is applied with a field strength of higher than approximately 9000 V/cm.

4. The method of claim 1, wherein the electrical pulse is applied with a field strength of higher than approximately 10,000 V/cm.

5. The method of claim 1, wherein the electrical pulse is applied with a shunt resistor of greater than approximately 100 OHM.