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AssigneeThe Regents of the University of California
InventorsRoger Y. Tsien, Gregor Zlokarnik
Estimating the term…
Substrates for β-lactamase of the general formula I 1 in which one of X and Y is a fluorescent donor moiety and the other is a quencher (which may or may not re-emit); R′ is selected from the group consisting of H, lower (i.e., alkyl of 1 to about 5 carbon atoms) and (CH2)nOH, in which n is 0 or an integer from 1 to 5; R″ is selected from the group consisting of H, physiologically acceptable metal and ammonium cations, —CHR2OCO(CH2)nCH3, —CHR2OCOC(CH3)3, acylthiomethyl, acyloxy-alpha-benzyl, delta-butyrolactonyl, methoxycarbonyloxymethyl, phenyl, methylsulphinylmethyl, beta-morpholinoethyl, dialkylaminoethyl, acyloxyalkyl, dialkylaminocarbonyloxymethyl and aliphatic, in which R2 is selected from the group consisting of H and lower alkyl; A is selected from the group consisting of S, O, SO, SO2 and CH2; and Z′ and Z″ are linkers for the fluorescent donor and quencher moieties. Methods of assaying β-lactamase activity and monitoring expression in systems using β-lactamase as a reporter gene also are disclosed.
A compound of the formula (I):
A recombinant nucleic acid molecule comprising expression control sequences adapted for function in a vertebrate cell and operably linked to a nucleotide sequence coding for the expression of a β-lactamase.
A recombinant nucleic acid molecule comprising expression control sequences adapted for function in a eukaryotic cell and operably linked to a nucleotide sequence coding for the expression of a cytosolic β-lactamase.
A mammalian host cell transfected with a recombinant nucleic acid molecule further comprising expression control sequences adapted for function in a mammalian cell and operably linked to the nucleotide sequence coding for the expression of a β-lactamase.
A method of determining the amount of β-lactamase activity in a cell comprising the steps of:
A method for monitoring the expression of a gene operably linked to a set of expression control sequences comprising:
A method for determining whether a test compound alters the expression of a gene operably linked to a set of expression control sequences comprising:
Dependencies are inferred from the claim text.