Human and mouse choline transporter cDNA
OwnerRandy D. Blakely, Subramaniam Apparsundaram +1inventors
InventorsRandy D. Blakely, Subramaniam Apparsundaram, Shawn Ferguson
Life of this patent
Estimating the term…
Abstract
Human high affinity choline transporter (CHT) cDNA was cloned from spinal cord and the primary structure and chromosomal localization have been determined. Mouse high affinity CHT cDNA was also cloned and characterized. An isolated polynucleotide, an isolated polypeptide, a recombinant host cell, a recombinant vector, a purified protein, an antibody, a nucleic acid detection kit, a method for screening cholinergic therapeutics, a method of treating a patient, a method of gene therapy and transgenic CHT mice are discussed.
Claims105
An isolated polynucleotide encoding a polypeptide comprising the amino sequence essentially as set forth in SEQ ID NO:
- 2.
A recombinant host cell comprising a DNA segment encoding an isolated human choline transporter.
A recombinant vector comprising a DNA segment encoding a human choline transporter polypeptide under the control of a promoter.
A purified and isolated polynucleotide wherein the polynucleotide comprises a sequence identical or complementary to between 14 and 100 contiguous nucleotides of SEQ ID NO:1.
A purified peptide comprising at least 10 contiguous amino acids of SEQ ID NO:2.
A method of using a DNA segment that encodes an isolated human choline transporter protein, comprising the steps of
- (a) preparing a recombinant vector in which a human choline transporter encoding said DNA segment is positioned under the control of a promoter;
- (b) introducing said recombinant vector into a host cell;
- (c) culturing said host cell under conditions effective to allow expression of the encoded protein or peptide;
- and
- (d) collecting said expressed protein or peptide.
An isolated polynucleotide encoding a polypeptide comprising the amino sequence essentially as set forth in SEQ ID NO:
- 4.
A recombinant host cell comprising a DNA segment encoding an isolated choline transporter having the amino acid sequence essentially as set forth in SEQ ID NO:4.
A recombinant vector comprising a DNA segment encoding a mouse choline transporter polypeptide under the control of a promoter.
A purified and isolated polynucleotide wherein the polynucleotide comprises a sequence identical or complementary to between 10 and 100 contiguous nucleotides of SEQ ID NO:3.
A purified peptide comprising at least 10 contiguous amino acids of SEQ ID NO:4.
An antibody that immunologically binds to a protein or peptide encoded by a contiguous sequence from the nucleic acid sequence essentially as set forth in SEQ ID NO:1.
An antibody that immunologically binds to a CHT protein or peptide that includes a contiguous amino acid sequence from SEQ ID NO:2.
A method of screening for cholinergic therapeutics comprising:
- (a) obtaining a candidate substance;
- (b) obtaining a recombinant cell comprising a polynucleotide encoding a choline transporter (CHT) polypeptide and a promoter heterologous to the polypeptide coding region, wherein said promoter directs expression of said CHT polypeptide;
- (c) combining candidate substance with said cell;
- and
- (d) determining whether said candidate substance modulates high-affinity choline uptake.
76 The method of claim 49, further comprising using said candidate substance to identify cholinergic neurons in a postmortem brain.
A method of treating a patient comprising:
- (a) obtaining a candidate substance;
- (b) obtaining a recombinant cell comprising a polynucleotide encoding a choline transporter (CHT) polypeptide and a promoter heterologous to the polypeptide coding region, wherein said promoter directs expression of said CHT polypeptide;
- (c) combining candidate substance with said cell;
- (d) determining whether said candidate substance modulates high-affinity choline uptake;
- and
- (e) delivering said candidate substance in a therapeutic formulation to a patient.
A nucleic acid detection kit comprising, in suitable container means, an isolated human choline transporter nucleic acid segment and a detection reagent.
A transgenic mouse, wherein said mouse lacks at least one functional mouse choline transporter (mCHT) allele.
A transgenic mouse, wherein the genome of said mouse comprises a choline transporter (CHT) encoding a DNA segment under the control of a heterologous promoter.
A transgenic mouse, wherein at least one mouse choline transporter (mCHT) allele is operably attacted to a detectable label.
A method comprising the step of delivering a polynucleotide encoding a choline transporter (CHT) polypeptide to a cell.
Dependencies are inferred from the claim text.