New light, oxygen, voltage domains, and proteins containing them, useful as fluorescent markers, e.g. in gene therapy, cloning and cell sorting, are altered to prevent covalent bonding to flavine mononucleotide

摘要

LOV (= light, oxygen, voltage) domains (A) in which at least one Cys is replaced by another amino acid (aa) that does not bind covalently to flavine mononucleotide are new. Independent claims are included for the following: (1) LOV-proteins (B) that include (A); (2) nucleic acid (NA) that encodes (A) or (B); (3) vector containing DNA Seq. (3), (4) or (5); (4) chromosome that contains at least one of Seq. (3)-(11) or (13); (5) genetically modified cells that contain a chromosome of (4) or vector of (3); (6) method for producing (A) or (B) by exchanging a Cys with an aa that can not form a covalent bond to FMN; (7) a protein (P) characterized by the sequence Gly-X-Asn-Cys-Arg-Phe-Leu-Gln (where X = any aa other than Cys) and having Cys at position 4 replaced by a different aa; and (8) FMN-carrying photoreceptor (PR) in which the FMN is not covalently bonded.