The United States of America As Represented by the Department of Health

The United States of America, As Represented by the Secretary, Department of Health and Human Services

David Goldman

Markku I Linnoila

Matti Virkkunen

Carl R Merril

Ullamari Pesonen

Markku Koulu

Chiara Mazzanti

Robert H Lipsky

David A Nielsen

Marc A Schuckit

Jaakko Lappalainen

Nakao Iwata

Sequence variations, such as polymorphisms, are detected by detecting differences between or among melting profiles of plural nucleic acids. Melting profiles are produced by observing the nucleic acids during a temperature change over a period of time. If the nucleic acids are diluted into solutions for analysis, the nucleic acid concentrations between or among the solutions can be substantially the same. The melting profiles and sequence variation can be used to identify changes in a wild-type nucleic acid sequence, such as a single-nucleotide polymorphism (SNP), a small insertion or deletion, or a small inversion.

Methods for detecting nucleic acid sequence variation

Compositions and methods based on a polymorphism in the gene encoding the &agr;6 subunit of the human GABA neurotransmitter receptor are disclosed. This polymorphism results in the substitution of a serine residue for a proline residue ordinarily present at amino acid position 385 of the &agr;6 polypeptide sequence. Significantly, the polymorphism was associated with decreased sensitivity to ethanol and benzodiazepine drugs.

Polynucleotides encoding polymorphic human GABAA receptor &agr;-6 subunit

An allelic variant of the 5HT7 serotonin receptor was discovered. Family study data indicate that this variant can be correlated to alchoholic offerders. DNA encoding the variant protein, the protein itself, vectors containing the variant gene and cell lines carrying a vector with the variant gene are part of the invention.

Allelic variation of the serotonin 5HT7 receptor

We describe identification of a non-conservative amino acid substitution in the 5-HT.sub.2C receptor gene. The variant was found using single strand conformational polymorphism (SSCP) analysis. By typing this polymorphism in CEPH families, the gene was genetically mapped on the long arm of the X chromosome. Since this polymorphism was not detectable as a conventional RFLP, we also developed a PCR-RFLP based method which allows rapid genotyping in populations and families.

Allelic variation of the serotonin 5HT.sub.2c receptor

A method of detecting abnormal serotonergic function in an impulsive human subject is disclosed. The method comprises detecting an L allele of a gene encoding tryptophan hydroxylase. The invention also includes an isolated nucleic acid having a sequence specific to an L allele of a gene encoding tryptophan hydroxylase. The nucleic acid may migrate in a denaturing gel at the same rate as a second nucleic acid specific to a corresponding region of a U allele of a gene encoding tryptophan hydroxylase, and migrate in a nondenaturing gel at a rate about 1.02 times the migration rate of the second nucleic acid.

Method for detecting abnormal serotonergic function

The invention provides an assay method for cellular and extracellular protein biodynamics comprising the use of radiochemical techniques in combination with high resolution protein separation procedures and recently developed silver stain techniques, to assess turnover of polypeptides over time. Quenching of polypeptide radiolables in silver stained gels is obviated by the use of .sup.14 C-labelled amino acids as precursors.

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Potomac, MD, United States of America

David Goldman