Ucp-2 promoter and use thereof
View Patent ↗This invention relates to a DNA containing uncoupling protein-2 (UCP-2) promoter region containing the regulator sequence, a transformant transformed with the DNA, a method for screening a compound or its salt that promotes or inhibits UCP-2 promoter activity characterized by use of the transformant, a method for screening an antiobestic drug, an antidiabetic drug, a depressor, an antihyperlipemic drug, and an antipyretic drug characterized by use of the transformant, a kit for screening a compound or its salt that promotes or inhibits UCP-2 promoter activity characterized by use of the transformant, and pharmaceutical composition containing a compound or its salt that promotes or inhibits UCP-2 promoter activity obtained using the screening method or the screening kit.
1. A method for screening for a compound or its salt that promotes or inhibits a human UCP-2 promoter activity, which comprises:
a. measuring the expression level of a reporter molecule in a transformant, with a DNA consisting of a human UCP-2 promoter sequence and a base sequence encoding the reporter molecule inserted downstream of the human UCP-2 promoter, contacted to a first sample of the compound or its salt and that in a control transformant, with a DNA consisting of a base sequence encoding the reporter molecule but with no DNA consisting of human UCP-2 promoter, contacted to a second sample of the compound or its salt,
wherein the human UCP-2 promoter sequence consists of (1) a base sequence consisting of nucleotides 1 to 2270 of SEQ ID NO: 1, or (2) a base sequence selected from the group consisting of nucleotides 255 to 430 of SEQ ID NO: 1, nucleotides 255 to 717 of SEQ ID NO: 1, nucleotides 717 to 1133 of SEQ ID NO: 1, nucleotides 1133 to 1389 of SEQ ID NO: 1, nucleotides 255 to 1857 of SEQ ID NO: 1, nucleotides 571 to 2270 of SEQ ID NO: 1, nucleotides 717 to 2270 of SEQ ID NO: 1, nucleotides 1133 to 2270 of SEQ ID NO: 1, nucleotides 1389 to 2270 of SEQ ID NO: 1, and nucleotides 1634 to 2270 of SEQ ID NO: 1; and
b. comparing the expression levels thereof.
2. A kit for screening for a compound or its salt that promotes or inhibits a human UCP-2 promoter activity, which comprises:
a. a medium for culturing a host animal cell line;
b. a plasmid for measurement of the human UCP-2 promoter activity, which comprises:
i. plasmid DNA carrying a DNA consisting of a human UCP-2 promoter sequence, wherein the human UCP-2 promoter sequence consists of (1) a base sequence consisting of nucleotides 1 to 2270 of SEQ ID NO: 1, or (2) a base sequence selected from the group consisting of nucleotides 255 to 430 of SEQ ID NO: 1, nucleotides 255 to 717 of SEQ ID NO: 1, nucleotides 717 to 1133 of SEQ ID NO: 1, nucleotides 1133 to 1389 of SEQ ID NO: 1, nucleotides 255 to 1857 of SEQ ID NO: 1, nucleotides 571 to 2270 of SEQ ID NO: 1, nucleotides 717 to 2270 of SEQ ID NO: 1, nucleotides 1133 to 2270 of SEQ ID NO: 1, nucleotides 1389 to 2270 of SEQ ID NO: 1, and nucleotides 1634 to 2270 of SEQ ID NO: 1; and
ii. a DNA consisting of a base sequence encoding a reporter molecule inserted downstream of the human UCP-2 promoter; and
c. a host animal cell line.
3. The kit of claim 2 , wherein the reporter molecule is a luciferase.