Isolated Photoprotein Aqdecay, and Its Use
The invention relates to the photoprotein AQdecay, to its nucleotide and amino acid sequences and to the activity and use of the photoprotein AQdecay.
1 . A nucleic acid molecule, or a functional fragment thereof, which is selected from the group consisting of:
a) a nucleic acid molecules which encodes a polypeptide which comprises the amino acid sequence disclosed by SEQ ID NO: 2;
b) a nucleic acid molecule which contains the sequence depicted by SEQ ID NO: 1;
c) a nucleic acid molecule whose complementary strands hybridize, under stringent conditions, with a nucleic acid molecule from a) or b) and whose expression products exhibit the biological function of a photoprotein; and
d) a nucleic acid molecules which differs from those mentioned under c) due to the degeneracy of the genetic code.
2 . A polypeptide, or a functional fragment thereof, which is encoded by a nucleic acid sequence according to claim 1 and possesses the property of a photoprotein.
3 . A photoprotein, or a functional fragment thereof, which possesses one or more mutations in positions 129 to 149 based on SEQ ID NO: 7 and which exhibits an altered chronological bioluminescence.
4 . A photoprotein, or a functional fragment thereof, which possesses a mutation in position 139 based on SEQ ID NO: 7 and which exhibits an altered chronological bioluminescence.
5 . A nucleic acid molecule which comprises a sequence which encodes a protein according to claims 3 and 4 .
6 . A nucleic acid molecule according to claim 1 or 5 which contains a functional promoter 5′ to the coding sequence.
7 . A recombinant DNA or RNA vector which contains a nucleic acid molecule according to claim 6 .
8 . A host cell which harbors a vector according to claim 7 .
9 . An oligonucleotide having more than 10 consecutive nucleotides which are identical with, or complementary to, a constituent sequence of a nucleic acid molecule according to claim 1 or 5 .
10 . A method for expressing the polypeptides according to claims 2 , 3 or 4 in a bacterial or eukaryotic cell or an in-vitro expression system.
11 . The method according to claim 10 further comprising the step of isolating the polypeptide.
12 . (canceled)
13 . (canceled)
14 . A method for preparing a photoprotein, comprising introducing one or more mutations in the region defined by positions 137 to 141 of SEQ ID NO: 7, thereby changing the chronological bioluminescence.
15 . A photoprotein, which is prepared by a method according to claim 14 .
16 . Use of a photoprotein according to claim 2 , 3 , 4 or 15 as a label or a reporter
17 . Use of a photoprotein according to claim 2 , 3 , 4 or 15 as a label or reporter in combination with other reporter genes.
18 . A variant of the photoprotein aequorin which exhibits an altered chronological bioluminescence.