β, β-carotene 15, 15′-monooxygenases, nucleic acid sequences coding therefor and their use
View Patent ↗Nucleic acid sequences coding for a protein having β,β-carotene 15,15′-monooxygenase activity and their uses in diagnostics, the synthesis of vitamin A and methods for the introduction of the β,β-carotene 15,15′-monooxygenase cDNA into host cells are disclosed.
1. An isolated nucleic acid sequence encoding a polypeptide having β,β-carotene 15,15′-monooxygenase activity comprising SEQ ID NO: 1 or a polypeptide having β,β-carotene 15,15′-monooxygenase activity and being at least 90% homologous to SEQ ID NO: 1 as determined by the Wisconsin Sequence Analysis Package GCG, Version 9.1 (1997).
2. An isolated nucleic acid sequence according to claim 1 , which comprises SEQ ID NO: 2 or a fragment thereof.
3. An isolated nucleic acid sequence according to claim 1 , wherein the nucleic acid is a deoxyribonucleic acid.
4. An isolated nucleic acid sequence comprising an antisense ribonucleic acid, which binds to the nucleic acid sequence according to claim 1 .
5. A primer for amplifying a polynucleotide coding for a polypeptide having β,β-carotene 15,15′-monooxygenase activity which primer consists essentially of a fragment of the nucleic acid sequence according to claim 1 .
6. A probe for detecting a polynucleotide coding for a polypeptide having β,β-carotene 15,15′-monooxygenase activity which probe consists essentially of a fragment of the nucleic acid sequence according to claim 1 .
7. A test kit for amplifying and/or detecting a polynucleotide or a fragment thereof coding for β,β-carotene 15,15′-monooxygenase wherein the test kit comprises at least one primer according to claim 5 .
8. A test kit for amplifying an or detecting a polynucleotide or a fragment thereof coding for β,β-carotene 15,15′-monooxygenase wherein the test kit comprises at least one probe to claim 6 .
9. A method for introducing β,β-carotene 15,15′-monooxygenase cDNA into a host cell comprising introducing a cDNA coding for a polypeptide having β,β-carotene 15,15′-monooxygenase activity comprising SEQ ID NO: 1 or a polypeptide having β,β-carotene 15,15′-monooxygenase activity and being at least 90% homologous to SEQ ID NO: 1 as determined by the Wisconsin Sequence Analysis Package GCG, Version 9.1 (1997) into a vector suitable for the host cell and introducing the vector into the host cell.
10. A method according to claim 9 wherein the host cell is a plant cell.
11. A method according to claim 9 wherein the host cell is a prokaryotic cell.
12. A method according to claim 9 wherein the host cell is a yeast cell or a fungal cell.
13. A method according to claim 9 wherein the host cell is an alga cell.
14. A method according to claim 9 wherein the host cell is a mammalian cell.
15. A method according to claim 14 wherein the mammalian cell is a human cell.
16. A host cell obtained by the method of claim 9 .
17. A host cell according to claim 16 which comprises a β,β-carotene 15,15′-monooxygenase cDNA obtained from another species.
18. A vector comprising a polynucleotide encoding a polypeptide having β,β-carotene 15,15′-monooxygenase activity comprising SEQ ID NO: 1 or a polypeptide having β,β-carotene 15,15′-monooxygenase activity and being at least 90% homologous to SEQ ID NO: 1 as determined by the Wisconsin Sequence Analysis Package GCG, Version 9.1 (1997).
19. A host cell transformed with the vector of claim 18 .