Invention:
Nucleotides 360-1890 from the human EP4 prostanoid receptor were PCR amplified from the small intestine, and digested with SspI and ScaI. The product was cloned into the plasmid ECoRV. This plasmid was digested with HindIII and XhoI, and the product was cloned into pCEP4, and transfected into 293-EBNA cells. Transformed lines were selected with hygromycin. The resulting protein was selected by PGE2 stimulation of cAMP formation. It was further characterized by immunofluorescence microscopy with a human EP2 receptor antibody and radioligand binding of [3H] prostaglandin E2.
Background:
University of Arizona researchers have created human embryonic kidney cells expressing the human EP4 prostanoid receptors. The new cell lines are expected to be a useful research tool for investigating receptor activity: they can be used in functional studies of receptor activation and second messenger signaling. They will also provide a sensitive and reliable measure of the potential for drugs to interact with the receptor.
Applications:
- Stably transformed cell lines
- Sensitive and reliable measure of the potential for drugs to interact with the receptors
Advantages:
- Useful in research into receptor activity and signaling
State of Development: Have stable cell lines expressing the receptors
