Codon-optimised nucleic acid coding for apoaequorin and uses thereof
OwnerJohn Andrew Printen, Wendy Irene Vernoninventors
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Abstract
The present invention relates to a codon-optimised nucleic acid sequence coding for apoaequorin polypeptide and uses thereof.
Claims13
A codon-optimised nucleic acid sequence coding for apoaequorin polypeptide.
An expression vector comprising a humanised apoaequorin gene and regulatory control sequences capable of directing expression of the humanised apoaequorin gene in a mammalian cell.
A recombinant host cell comprising a humanised apoaequorin gene.
A method for producing apoaequorin protein comprising the steps of:
- (i) preparing a recombinant expression vector in which a humanised apoaequorin gene is positioned under the regulatory control of a promoter operative in a mammalian host cell;
- (ii) introducing said recombinant expression vector into a suitable mammalian host cell;
- (iii) culturing the host cell under conditions suitable for allowing expression of the encoded apoaequorin protein;
- and optionally, purifying said expressed apoaequorin protein from a significant amount of other cellular proteins.
A method of increasing the magnitude of aequorin luminescence comprising, introducing into a host cell nucleic acid comprising a codon-optimised nucleic acid sequence coding for apoaequorin polypeptide operably linked to regulatory sequences capable of effecting expression of the codon-optimised nucleic acid to produce said apoaequorin polypeptide.
The use of a codon-optimised apoaequorin nucleic acid sequence for enhancing the magnitude of aequorin luminescence in a host cell.
A method for measuring the ability of a compound to inhibit a receptor which mediates changes in intracellular calcium flux when activated, comprising contacting a host cell engineered to express codon optimised apoaequorin with the luciferin coelenterazine and measuring the amount of luminescence produced with or without addition of a test compound.
Dependencies are inferred from the claim text.