Novel clock gene promoter
The present invention discloses a Period2 gene promoter, a construct containing the promoter and a reporter gene, a cell containing the construct, a transgenic animal harboring the construct, and a method for screening a substance which controls expression or oscillatory expression of a biological clock gene. The screening method uses the above cell, a suprachiasmatic nucleus section or peripheral tissue of the above transgenic animal or the above transgenic animal.
1 - 10 . (canceled)
11 . An isolated DNA which maintains a basal promoter activity and has a promoter activity transcriptionally-activated by a BMAL1/CLOCK heterodimer, which comprises the nucleotide sequence consisting of nucleotides at positions 3,820 to 6,068 in the nucleotide sequence represented by SEQ ID NO:2.
12 . The DNA according to claim 11 , which consists of the nucleotide sequence of nucleotides at positions 3,820 to 6,068 in the nucleotide sequence represented by SEQ ID NO:2.
13 . A construct which comprises the DNA according to claim 11 or 12 operably linked to a reporter gene.
14 . A cell which comprises the construct according to claim 13 .
15 . A method for screening a substance which controls expression of Period2 gene, comprising the steps of:
allowing the cell according to claim 14 to contact with a substance to be tested, and
measuring activity of the reporter gene.
16 . A transgenic rat or mouse transfected with the construct according to claim 13 , and wherein the suprachiasmatic nucleus and/or peripheral tissues of the rat or mouse exhibit the function of reporter gene expression.
17 . The transgenic rat according to claim 16 .
18 . The transgenic rat or mouse according to claim 16 , wherein the reporter gene is one member selected from the group consisting of a gene encoding luciferase, a gene encoding secretion type alkaline phosphatase (SEAP), a gene encoding green fluorescent protein (GFP), a gene encoding chloramphenical acetyltransferase (CAT), a gene encoding β-glucuronidase (GUS), a gene encoding β-D-galactosidase and a gene encoding aequorin.
19 . The transgenic rat according to claim 17 , wherein the reporter gene is one member selected from the group consisting of a gene encoding luciferase, a gene encoding secretion type alkaline phosphatase (SEAP), a gene encoding green fluorescent protein (GFP), a gene encoding chloramphenical acetyltransferase (CAT), a gene encoding β-glucuronidase (GUS), a gene encoding β-D-galactosidase and a gene encoding aequorin.
20 . A method for screening a substance which controls expression and/or oscillatory expression of Period2 gene, comprising the steps of:
allowing the cell according to claim 14 to react with a substance to be tested, and
measuring activity of the reporter gene for oscillatory expression of the Period2 gene.
21 . A method for screening a substance which controls expression and/or oscillatory expression of Period2 gene, comprising the steps of:
administering a substance to be tested to the transgenic rat or mouse according to claim 16 , and
measuring activity of the reporter gene in the suprachiasmatic nucleus of the animal for oscillatory expression of the Period2 gene.
22 . A method for screening a substance which controls expression and/or oscillatory expression of Period2 gene, comprising the steps of:
allowing a suprachiasmatic nucleus section or peripheral tissue of the transgenic rat or mouse according to claim 16 to react with a substance to be tested, and
measuring activity of the reporter gene for oscillatory expression of the Period2 gene.