Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 1 de 1
Filter
Add more filters










Database
Language
Publication year range
1.
Proc Natl Acad Sci U S A ; 103(7): 2052-6, 2006 Feb 14.
Article in English | MEDLINE | ID: mdl-16461889

ABSTRACT

Fluorescent proteins have proven to be excellent reporters and biochemical sensors with a wide range of applications. In a split form, they are not fluorescent, but their fluorescence can be restored by supplementary protein-protein or protein-nucleic acid interactions that reassemble the split polypeptides. However, in prior studies, it took hours to restore the fluorescence of a split fluorescent protein because the formation of the protein chromophore slowly occurred de novo concurrently with reassembly. Here we provide evidence that a fluorogenic chromophore can self-catalytically form within an isolated N-terminal fragment of the enhanced green fluorescent protein (EGFP). We show that restoration of the split protein fluorescence can be driven by nucleic acid complementary interactions. In our assay, fluorescence development is fast (within a few minutes) when complementary oligonucleotide-linked fragments of the split EGFP are combined. The ability of our EGFP system to respond quickly to DNA hybridization should be useful for detecting the kinetics of many other types of pairwise interactions both in vitro and in living cells.


Subject(s)
DNA/chemistry , Green Fluorescent Proteins/chemistry , Green Fluorescent Proteins/genetics , Spectrometry, Fluorescence/methods , Fluorescence , Nucleic Acid Hybridization , Oligonucleotides/chemistry , Protein Folding , Sequence Deletion
SELECTION OF CITATIONS
SEARCH DETAIL
...