Method for testing a therapeutic or preventive agent for hyperlipidemia
First Claim
1. A method for testing an effect of a substance as a therapeutic or preventive agent for hyperlipidemia comprising:
- (i) culturing cells in the presence or absence of a test substance;
(ii) detecting the amount of expression of a mRNA which has a nucleotide sequence of one of the following sequences (a) to (e), wherein t in the sequence is read as u, in the cultured cells obtained in step (i);
(a) a nucleotide sequence having the nucleotide numbers 47-1411 of SEQ ID No. 1 of the Sequence Listing;
(b) a nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing;
(c) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pBK/m55-1 SANK72199 (FERM BP-6940);
(d) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pTrip/h55-1 SANK72299 (FERM BP-6941);
(e) a nucleotide sequence which hybridizes with the polynucleotide consisting essentially of an antisense sequence of the nucleotide sequence of one of the nucleotide sequences (a) to (d) under stringent conditions, and encodes a polypeptide having an activity of raising neutral fat concentration in blood; and
(iii) comparing the amount of expression of mRNA between the cells cultured in the absence of the substance and the cells cultured in the presence of the substance, as a result of detection step (ii).
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Abstract
A method for testing a therapeutic or preventive agent for hyperlipidemia, and a nucleic acid probe, a primer, and an antibody which are used in the method. Specifically, a method for testing the effect of a test substance as a therapeutic or preventive agent for hyperlipidemia by expression of a gene having a sequence as set forth in SEQ ID No. 1 or SEQ ID No. 2 of the Sequence Listing which participates in regulation of neutral-fat concentration in the blood of a mammal, and a nucleic acid probe, primer or antibody used for the method. The present invention makes it possible to search for a candidate substance for a therapeutic or preventive agent for hyperlipidemia.
14 Citations
56 Claims
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1. A method for testing an effect of a substance as a therapeutic or preventive agent for hyperlipidemia comprising:
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(i) culturing cells in the presence or absence of a test substance;
(ii) detecting the amount of expression of a mRNA which has a nucleotide sequence of one of the following sequences (a) to (e), wherein t in the sequence is read as u, in the cultured cells obtained in step (i);
(a) a nucleotide sequence having the nucleotide numbers 47-1411 of SEQ ID No. 1 of the Sequence Listing;
(b) a nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing;
(c) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pBK/m55-1 SANK72199 (FERM BP-6940);
(d) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pTrip/h55-1 SANK72299 (FERM BP-6941);
(e) a nucleotide sequence which hybridizes with the polynucleotide consisting essentially of an antisense sequence of the nucleotide sequence of one of the nucleotide sequences (a) to (d) under stringent conditions, and encodes a polypeptide having an activity of raising neutral fat concentration in blood; and
(iii) comparing the amount of expression of mRNA between the cells cultured in the absence of the substance and the cells cultured in the presence of the substance, as a result of detection step (ii). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30)
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31. A polynucleotide having a nucleotide sequence which hybridizes to a mRNA containing a nucleotide sequence selected from the following sequences (a) to (d), wherein t in the sequence is read as u, under stringent conditions, excluding nucleotide sequences in the following (a) to (d):
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(a) a nucleotide sequence having the nucleotide numbers 47-1411 of SEQ ID No. 1 of the Sequence Listing;
(b) a nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing;
(c) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pBK/m55-1 SANK72199 (FERM BP-6940);
(d) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pTrip/h55-1 SANK72299 (FERM BP-6941).
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32. A DNA consisting essentially of at least 15 nucleotides of the nucleotide sequence having the nucleotide numbers 47-1411 of SEQ ID No. 1 of the Sequence Listing wherein one or more of nucleotides are deleted at one end or both ends thereof.
- 33. A DNA consisting essentially of at least 15 nucleotides of the nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing wherein one or more of nucleotides are deleted at one end or both ends.
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34. A method for testing the effect of a substance as a therapeutic or preventive agent for hyperlipidemia comprising:
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(i) culturing cells in the presence or absence of a test substance;
(ii) detecting the amount of production of polypeptide having an amino acid sequence encoded by the nucleotide sequence of one of the following sequences (a) to (e) or a part thereof in a supernatant of the cultured cells obtained in step (i) using an antibody specifically recognizing the polypeptide;
(a) a nucleotide sequence having the nucleotide numbers 47-1411 of SEQ ID No. 1 of the Sequence Listing;
(b) a nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing;
(c) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pBK/m55-1 SANK72199 (FERM BP-6940);
(d) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pTrip/h55-1 SANK72299 (FERM BP-6941);
(e) a nucleotide sequence which hybridizes with a polynucleotide consisting essentially of an antisense sequence of a nucleotide sequence in one of the sequences (a) to (d) under stringent conditions, and encodes a polypeptide having an activity of raising neutral fat concentration in blood; and
(iii) comparing the amount of production of the polypeptide between the cells cultured in the absence of the substance and the cells cultured in the presence of the substance, as a result of that detected in step (ii).
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46. An antibody which specifically recognizes a polypeptide consisting of the amino acid sequence encoded by one of the following sequences (a) to (e) or a part thereof:
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(a) the nucleotide sequence having the nucleotide numbers 47-1411 of SEQ ID No. 1 of the Sequence Listing;
(b) the nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing;
(c) the nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pBK/m55-1 SANK72199 (FERM BP-6940);
(d) the nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pTrip/h55-1 SANK72299 (FERM BP-6941);
(e) the nucleotide sequence which hybridizes with a polynucleotide consisting essentially of an antisense sequence of a nucleotide sequence in one of the nucleotide sequences (a) to (d) under stringent conditions, and encodes a polypeptide having an activity of raising neutral fat concentration in blood.
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50. A method for testing the effect of a substance as a therapeutic or preventive agent for hyperlipidemia comprising:
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(i) administrating the substance to be tested to an animal other than a human obtained by genetic manipulation in which a foreign gene containing the nucleotide sequence of one of the following sequences (a) to (e) is introduced so that the expression of the gene can be detected;
(a) a nucleotide sequence having the nucleotide numbers 47-1411 of SEQ ID No. 1 of the Sequence Listing;
(b) a nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing;
(c) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pBK/m55-1 SANK72199 (FERM BP-6940);
(d) a nucleotide sequence of DNA incorporated into a phagemid carried by transformed Escherichia coli E.coli pTrip/h55-1 SANK72299 (FERM BP-6941);
(e) a nucleotide sequence which hybridizes with a polynucleotide consisting essentially of an antisense sequence of the nucleotide sequence of one of the nucleotide sequences (a) to (d) under stringent conditions, and encodes a polypeptide having an activity of raising neutral fat concentration in blood; and
(ii) measuring the concentration of neutral-fat in the blood of the animal in step (i). - View Dependent Claims (51, 52)
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53. An antibody for testing a therapeutic or preventive agent for hyperlipidemia which specifically recognizes a polypeptide consisting essentially of the amino acid sequence having the amino acid numbers 17-455 of SEQ ID No. 2 of the Sequence Listing or a part thereof, a polypeptide which consists essentially of the amino acid sequence having the amino acid numbers 19-455 of SEQ ID No. 2 of the Sequence Listing or a part thereof, or a polypeptide which consists essentially of the amino acid sequence having the amino acid numbers 17-460 of SEQ ID No. 4 of the Sequence Listing or a part thereof.
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54. In a test kit for a therapeutic or preventive agent for hyperlipidemia which includes an antibody, wherein the improvement comprises the antibody being an antibody that specifically recognizes a polypeptide consisting essentially of the amino acid sequence having the amino acid numbers 17-455 of SEQ ID No. 2 of the Sequence Listing or a part thereof, a polypeptide which consists essentially of the amino acid sequence having the amino acid numbers 19-455 of SEQ ID No. 2 of the Sequence Listing or a part thereof, or a polypeptide which consists essentially of the amino acid sequence having the amino acid numbers 17-460 of SEQ ID No. 4 of the Sequence Listing or a part thereof.
- 55. A DNA or RNA consisting essentially of an antisense sequence of the nucleotide sequence which consists essentially of a continuous 15 or 30 nucleotides in the nucleotide sequence having the nucleotide numbers 78-1457 of SEQ ID No. 3 of the Sequence Listing.
Specification