5792846 : Human calcium channel compositions and methods
21 CLAIMS
What is claimed is:
1. An isolated DNA molecule, comprising a sequence of nucleotides that encodes an .alpha.1B -subunit of human
calcium channel, wherein:
the sequence of nucleotides encoding the .alpha.1B -subunit is selected from the group consisting of:
(a) a sequence of nucleotides that encodes a naturally-occurring human .alpha.1B -subunit and comprises
the sequence of nucleotides set forth in SEQ ID No. 7 or 8;
(b) a sequence of nucleotides that encodes an .alpha.1B -subunit, wherein:
the DNA molecule comprising this sequence hybridizes under conditions of high stringency to DNA
that if fully complementary to an mRNA transcript native to a human cell; and
the transcript: (i) encodes an .alpha.1B -subunit, (ii) comprises the sequence of nucleotides set forth
in SEQ ID No. 8 or 7 in which T residues are replaced by U, and (iii) hybridizes under conditions of
high stringency to any probe containing at least about 14 contiguous bases from the coding portion
the sequence of nucleotides that encodes the .alpha.1B -subunit;
(c) a sequence of nucleotides that encodes an .alpha.1 -subunit that comprises the sequence of amino acids
of the .alpha.1 -subunit set forth in SEQ ID NO. 47 or SEQ ID NO. 48; and
(d) a sequence of nucleotides comprising degenerate codons with sequence of (b).
2. A cultured eukaryotic cell, comprising heterologous DNA that encodes a human calcium channel .alpha.1 -subunit
encoded by the DNA of claim 1, wherein the cell is a mammalian cell or yeast cell. (snip)
15. An isolated eukaryotic cell with a functional, heterologous calcium channel, produced by a process, comprising:
introducing into the cell RNA into that is translatable in the cell to afford an .alpha.1 -subunit of a human calcium
channel, wherein the .alpha.1 -subunit comprises an amino acid sequence encoded by the DNA molecule of claim
1.
16. An isolated eukaryotic cell with a functional, heterologous calcium channel, produced by a process, comprising:
introducing into the cell at least one RNA transcript selected from the group consisting of a first RNA that is
translatable in said cell into an .alpha.1B -subunit of a human calcium channel, a second RNA which is translatable
in said cell into a .beta.-subunit of a human calcium channel, a third RNA which is translatable in said cell into an
.alpha.2 -subunit of a human calcium channel, and a fourth RNA which is translatable in said cell into a
.gamma.-subunit of a human calcium channel, wherein the cell is a mammalian cell, yeast cell or amphibian oocyte,
wherein:
the .alpha.1 -subunit comprises a sequence of amino acids encoded by the DNA molecule of claim 1;
the .alpha.2 -subunit is a human calcium channel .alpha.2 -subunit that has an amino acid sequence set forth
in SEQ ID NO. 53, 54, 55 or 56, or is an .alpha.2a, .alpha.2c, .alpha.2d, or an .alpha.2e subtype and is
encoded by DNA that is capable of hybridizing under conditions of high stringency with DNA that is fully
complementary to an mRNA transcript present in a human cell that encodes one of the aforesaid human
calcium channel .alpha.2 -subunits having an amino acid sequence set forth in any of the SEQ ID NOs. 53,
54, 55 or 56, such that any probe that contains at least 14 contiguous bases from the coding portion of the
DNA that encodes the .alpha.2 -subunit hybridizes under conditions of high stringency to the DNA that is
fully complementary to the mRNA transcript present in a human cell;
the .beta.-subunit is selected from the group consisting of
a .beta.-subunit comprising the amino acids set forth in SEQ ID NO. 40, a .beta.-subunit comprising
the amino acids set forth in SEQ ID NO. 41, a .beta.-subunit comprising the amino acids set forth in
SEQ ID NO. 42, a .beta.-subunit comprising the amino acids set forth in SEQ ID NO. 43, a
.beta.-subunit comprising the amino acids set forth in SEQ ID NO. 44, and a human calcium channel
.beta.-subunit, wherein the .beta.-subunit is encoded by DNA capable of hybridizing under
conditions of high stringency with DNA that is fully complementary to an mRNA transcript present in
a human cell that encodes one of the aforesaid .beta.-subunits such that any probe that contains at
least 14 contiguous bases from the coding portion of the DNA that encodes the .beta.-subunit
hybridizes under conditions of high stringency to the DNA that is fully complementary to the mRNA
transcript present in a human cell that encodes any of the aforesaid .beta.-subunits;
the .gamma.-subunit comprises amino acids set forth in SEQ ID No. 57, or is a human calcium channel
.gamma.-subunit encoded by DNA capable of hybridizing under conditions of high stringency with DNA
that is complementary to an mRNA transcript present in a human cell that comprises the sequence of
nucleotides set forth in SEQ ID NO. 14 such that any probe that contains at least 14 contiguous bases
from the coding portion of the DNA that encodes the .gamma.-subunit hybridizes under conditions of high
stringency to the DNA that is fully complementary to the mRNA transcript present in a human cell; and
the only heterologous ion channels that are expressed by the cell are calcium channels.
17. An isolated eukaryotic cell with a functional, heterologous calcium channel, produced by a process, comprising
introducing into the cell at least one RNA transcript selected from the group consisting of a first RNA that is translatable
in said cell into an .alpha.1B -subunit of a human calcium channel, a second RNA which is translatable in said cell into a
.beta.-subunit of a human calcium channel, a third RNA which is translatable in said cell into an .alpha.2b -subunit of a
human calcium channel, and a fourth RNA which is translatable in said cell into a .gamma.-subunit of a human calcium
channel, wherein the cell is a mammalian cell, yeast cell or amphibian oocyte, wherein:
the .alpha.1 -subunit comprises a sequence of amino acids encoded by the DNA molecule of claim 52;
the .alpha.2b -subunit is a human calcium channel .alpha.2 -subunit that has an amino acid sequence set forth in
SEQ ID NO. 52 or is encoded by DNA that is capable of hybridizing under conditions of high stringency with
DNA that is fully complementary to an mRNA transcript native to a human cell that encodes the aforesaid human
calcium channel .alpha.2b subunit, such that any probe that contains at least 14 contiguous bases from the coding
portion of the DNA that encodes the .alpha.2b -subunit hybridizes under conditions of high stringency to the DNA
that is fully complementary to the mRNA transcript native to a human cell that encodes any of the aforesaid
subunits; and
the .beta.-subunit is selected from the group consisting of
a .beta.-subunit comprising the amino acids set forth in SEQ ID NO. 40, a .beta.-subunit comprising the
amino acids set forth in SEQ ID NO. 41, a .beta.-subunit comprising the amino acids set forth in SEQ ID
NO. 42, a .beta.-subunit comprising the amino acids set forth in SEQ ID NO. 43, a .beta.-subunit
comprising the amino acids set forth in SEQ ID NO. 44, and a human calcium channel .beta.-subunit,
wherein the .beta.-subunit is encoded by DNA capable of hybridizing under conditions of high stringency
with DNA that is fully complementary to an mRNA transcript present in a human cell that encodes one of
the aforesaid .beta.-subunits such that any probe that contains at least 14 contiguous bases from the coding
portion of the DNA that encodes the .beta.-subunit hybridizes under conditions of high stringency to the
DNA that is fully complementary to the mRNA transcript present in a human cell that encodes any of the
aforesaid .beta.-subunits;
the .gamma.-subunit comprises amino acids set forth in SEQ ID No. 57, or is a human calcium channel
.gamma.-subunit encoded by DNA capable of hybridizing under conditions of high stringency with DNA that is
complementary to an mRNA transcript present in a human cell that comprises the sequence of nucleotides set
forth in SEQ ID NO. 14 such that any probe that contains at least 14 contiguous bases from the coding portion of
the DNA that encodes the .gamma.-subunit hybridizes under conditions of high stringency to the DNA that is fully
complementary to the mRNA transcript present in a human cell; and
the only heterologous ion channels that are expressed by the cell are calcium channels. (snip) |
