Transformation of algal cells
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.
1. A method for transforming an algal cell, the method comprising:
preparing a transformation construct, the transformation construct including a gene involved in lipid biosynthesis;
preparing a particle for bombarding the algal cell;
adhering the transformation construct to the particle;
bombarding the algal cell with the particle;
growing the algal cell into a colony;
wherein the particle is a gold particle smaller than or equal to approximately 1 micrometer average diameter; and
wherein the preparing of the gold particle includes sonicating the gold particle with ultrasound prior to the adhering of the transformation construct to the gold particle.
2. The method of claim 1 , wherein the transformation construct is replicated within a nuclear genome of the algal cell.
3. The method of claim 1 , wherein the growing of the algal cell is in a nutrient medium.
4. The method of claim 1 , wherein the transformation construct is a nucleic acid in an expression cassette.
5. The method of claim 4 , wherein the expression cassette further comprises:
a promoter functional in the algal cell operably linked to the gene involved in lipid biosynthesis;
a selectable transformation marker gene; and
a termination region functional in the algal cell operably linked to the gene involved in lipid biosynthesis.
6. The method of claim 5 , wherein the selectable transformation marker gene is a gene capable of conferring antibiotic resistance to the algal cell.
7. The method of claim 6 , the method further comprising:
selecting the algal cell growing in a selective medium having an antibiotic the gene has conferred resistance upon to the algal cell.
8. The method of claim 1 , wherein the algal cell is an auxotrophic mutant.
9. The method of claim 1 , wherein the transformation construct includes a sh ble gene.
10. The method of claim 1 , wherein the transformation construct includes a hygromycin B phosphotransferase gene.
11. The method of claim 1 , wherein the transformation construct includes a nitrate reductase gene.
12. The method of claim 1 , wherein the transformation construct includes a blastocidin S deaminase gene.
13. The method of claim 1 , wherein the adhering of the transformation construct to the particle includes precipitating DNA of the transformation construct to the particle.