Aurora Algae, Inc.

Aurora Alage, Inc.

Oliver Kilian

Bertrand Vick

Provided herein are exemplary isolated nucleotide sequences encoding polypeptides having elongase activity, which utilize fatty acids as substrates.

Algal elongase 6

Isolated nucleotide sequences encoding a promoter of the Acyl Carrier Protein ('ACP').

ACP promoter

Exemplary transformation methods are provided for introducing deoxyribonucleic acid (DNA) into the nucleus of an algal cell. A transformation construct may be prepared, with the transformation construct having a first sequence of DNA similar to a corresponding first sequence of nuclear DNA, a second sequence of DNA similar to a corresponding second sequence of the nuclear DNA, and a sequence of DNA of interest inserted between the first and second sequences of DNA of the transformation construct. A target sequence of DNA inserted between the first and second corresponding sequences of the nuclear DNA may be transformed, resulting in replacement of the target sequence of DNA with the sequence of DNA of interest. Also provided are exemplary transformation constructs, with some transformation constructs having a first sequence of DNA similar to a corresponding first sequence of nuclear DNA of an algal cell, a second sequence of DNA similar to a corresponding second sequence of nuclear DNA of the algal cell, and a sequence of DNA of interest inserted between the first and second sequences of the transformation construct.

Homologous recombination in an algal nuclear genome

Algal elongases

Provided herein are exemplary isolated nucleotide sequences encoding polypeptides having desaturase activity, which utilize fatty acids as substrates.

Algal desaturases

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.

Transformation of algal cells

Provided herein are exemplary vectors for transforming algal cells. In exemplary embodiments, the vector comprises a Violaxanthin-chlorophyll a binding protein (Vcp) promoter driving expression of an antibiotic resistance gene in an algal cell. Embodiments of the invention may be used to introduce a gene (or genes) into the alga, such that the gene(s) are expressed and functional. This unprecedented ability to transformwith high efficiency makes possible new developments in phycology, aquaculture and biofuels applications.

VCP-based vectors for algal cell transformation

Provided herein are exemplary genes, constructs and methods for the formation of triacylglycerols (TAGs). The exemplary genes include a phosphatic acid phosphohydrolase (PA Hydrolase) gene, a diacylglycerol o-acyltransferase (DAGAT2A) gene, and a phospholipid:diacylglycerol acyltransferase (LROI) gene.

Genes for enhanced lipid metabolism for accumulation of lipids

Provided herein are exemplary vectors for transforming algal cells. In exemplary embodiments, the vector comprises a Violaxanthin-chlorophyll a binding protein (Vcp) promoter driving expression of an antibiotic resistance gene in an algal cell. Embodiments of the invention may be used to introduce a gene (or genes) into the algasuch that the gene(s) are expressed and functional. This unprecedented ability to transformwith high efficiency makes possible new developments in phycology, aquaculture and biofuels applications.

Provided herein are exemplary methods for production of biomass with a cyanobacterial isolate having auto-flocculation properties. One exemplary method includes isolating a cyanobacterial strain having a 16S ribosomal RNA sequence corresponding to SEQ. ID. NO. 1 herein, inoculating an algae cultivation system with the cyanobacterial strain, growing the cyanobacterial strain, and harvesting the biomass produced by the cyanobacterial strain. According to a further method, the harvesting of the biomass comprises ceasing agitation of the algae cultivation system, and pooling a slurry of the biomass produced by the cyanobacterial strain. In a further method, the harvesting of the biomass may comprise ceasing agitation within the algae cultivation system and/or allowing the biomass produced by the cyanobacterial strain to settle to near or at a bottom of the algae cultivation system. Also provided herein are exemplary cyanobacterial strains having flocculation properties for production of a biomass.

Cyanobacterial isolates having auto-flocculation and settling properties

Exemplary embodiments provided herein include novel promoters isolated from the microalgae,. These promoters drive gene expression in a bidirectional manner, and are especially useful for the genetic manipulation ofand other organisms. The inventors herein successfully used these promoters (in both parallel and antiparallel orientations with respect to a Sh ble gene cassette) to impart zeocine-resistance to

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Alameda, CA, United States of America

Oliver Kilian