DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS In a preferred aspect of this invention, the invention relates to a method for the detection of an analyte in a sample.
The method employs DNA, preferably a DNA, that comprises a capturing agent positioned on the DNA, at least one regulatory region, such as a promoter, capable of promoting transcription from a reporter, and a reporter capable of being transcribed to RNA when the promoter is activated.
Preferably, the DNA also includes an adaptor nucleotide sequence, wherein at least a portion converts one restriction endonuclease site to another, a portion that encodes an RNA transcript signal (e.
g.
, a poly A tail or a TATA signal), and, optionally, a portion that encodes a promoter.
Examples include SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4.
SEQ ID NO: 1 and SEQ ID NO:2 each include a portion that converts an Xba I restriction site to a BamHI and a portion that encodes a poly A tail.
SEQ ID NO:3, and SEQ ID NO:4 each include a portion that converts a Hind III restriction site to an Eco RI site, a portion that includes a TATA signal, and a portion that encodes an Elb minimal promoter.
Other adaptor nucleotide sequences can be designed by those skilled in the art.
The assay further comprises an activator complex.
The activator complex includes an activator that is capable of facilitating RNA polymerase-mediated transcription from the reporter in the presence of a suitable promoter.
The activator complex also includes an analyte recognizing region that is capable of specifically recognizing the analyte in the assay.
Preferred transcriptional activators are gal4 (Fields S and Song O-K, Nature 340:245-6, 1989), ZEBRA (Chi T.
and Carey M.
, Mol.
and Cell.
BioL 13:7045-55, 1993), and VP16 (Tantin D.
et al.
Methods in Enzymology.
274:133-149, 1996) for eukaryotic-based systems and .
lambda.
cI-CTD for a prokaryotic system (Dove SL et al.
Nature 386:627-30, 1997), such as a system using a prokaryotic promoter including those found in bacteria such as E coli
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