The Salk Institute for Biological Studies

Other

James Warner Patrick

Stephen Fox Heinemann

Evan Samuel Deneris

Robert M Duvoisin

Marc Charles Ballivet

James Richard Boulter

Keiji Wada

Eden Deer Heinemann

Daniel Jay Goldman

John Gerard Connolly

Barbara D Boss

W Maxwell Cowan

Leslie E Orgel

Robert Michael Duvoisin

The present invention relates to a family of neuronal nicotinic acetylcholine receptors comprised of neuronal agonist and non-agonist binding subunits, and DNA sequences encoding such subunits. These novel neuronal nicotinic acetylcholine receptor subunits include the agonist binding subunits alpha2, alpha3, alpha4, and alpha5, plus non-agonist binding subunits beta2, beta3 and beta4. Representative cDNA clones that contain the DNA sequences of the invention have been deposited with the American Type Culture Collection for patent purposes.

Neuronal nicotinic acetylcholine receptor compositions

Neuronal nicotinic acetylcholihne receptor compositions

The present invention discloses a new neuronal nicotinic acetylcholine receptor subunit, .beta.4. The new subunit can form functional combinations with other neuronal nicotinic acetylcholine receptor subunits, including, but not limited to, alpha2, alpha3, alpha4 and beta2. A cDNA clone containing the DNA sequences that encode the novel receptor subunit of the invention has been deposited with the American Type Culture Collection for patent purposes.

Recombinantly produced .beta.4 neuronal nicotinic acetylcholine receptor

Neuronal nicotinic acetylcholine receptor assay

DNA encoding neuronal nicotinic acetylcholine receptor compositions

A sequence in the coat glycoprotein of rabies virus is identified as the molecular basis for an essential step in the pathogenesis of the virus, the binding of virus to acetylcholine receptor at neuromuscular junctions prior to virus uptake into peripheral nerves. Based on this discovery, synthetic peptide-based, anti-rabies vaccines are prepared. The active ingredient of such vaccines is a synthetic protein, which is a conjugate with an immunogenic carrier protein of a synthetic peptide with a sequence which includes a sequence which is substantially the same as a substantial portion of the sequence of the acetylcholine receptor-binding segment of the rabies virus coat protein. Anti-rabies antisera, and anti-rabies antibodies, are prepared by injecting a mammal with a synthetic protein of the invention in a manner that induces an immune response in the mammal against the synthetic protein. Hybridomas which secrete anti-rabies antibodies, which bind to specific epitopes on or near the acetylcholine receptor binding segment on the rabies virus coat glycoprotein, are prepared with B cells immunized in vitro with a synthetic peptide or synthetic protein of the invention or taken from a mammal inoculated with such a peptide or protein. The anti-rabies vaccines of the invention avoid risks associated with currently available anti-rabies vaccines. The antisera and antibodies of the invention are useful therapeutically to induce passive anti-rabies immunity in a mammal that has been exposed to rabies virus. The antisera and antibodies are also useful for diagnosing whether a mammal is rabid and whether a mammal has been exposed to live rabies virus.

Synthetic peptide-based anti-rabies compositions and methods

A first mixture is prepared that contains labeled chain fragments which each has a common end adjacent to a primary nucleotide and a termination at a position from the primary through an nth nucleotide, the first mixture containing nucleotide chain fragments of each length from termination at the primary through termination of the nth nucleotide. A second mixture is prepared that contains labeled chain fragments beginning at the common end and terminating at positions from the first through the nth nucleotide, the second mixture containing chain fragments of each length terminating wherever either a first or a second of the four nucleotides occurs. A third mixture is prepared that contains labeled chain fragments beginning at the common end and terminating at a position from the first through the nth nucleotide, the third mixture containing chain fragments of each length terminating wherever the first or a third of the four nucleotide sequences occurs. The chains are electrophoresed with the first mixture as the center lane. This three-lane system provides a unique band pattern for each of the four nucleotides and permits the sequence to be read merely by directly comparing each of the flanking lanes with the fully stepped center lane. This system has important advantages in reducing reading errors, particularly when read with computer-assisted scanning apparatus.

DNA sequencing

Growing community of inventors

Houston, TX, United States of America

James Warner Patrick