| 摘要 |
PROBLEM TO BE SOLVED: To obtain a new RNA containing pyrene-incorporated base(s) in its constituent base sequence, and emitting intense fluorescence through its hybridization with the corresponding complementary strand, therefore assayable in its reaction in a homogenous system by the aid of measuring the above fluorescence. SOLUTION: This new RNA is such one as to contain pyrene-incorporated base(s) in its constituent base sequence, emit intense fluorescence when hybridized with the corresponding complementary strand, and therefore be assayable in a homogeneous system. The assay technique is useful in e.g. in-situ detection, variant detection. This pyrene-modified RNA is obtained according to the following procedure: a reaction is carried out between uridine with the hydroxyl groups on 3'- and 5'-sites protected with trityl groups and 1-chloromethylpyrene followed by deprotection to covert into 2'-O-(1-pyrenylmethyl)uridine, with which a 4,4'-dimethoxytritylpyrene-modified uridine-phosphorodiamidate reagent is prepared, and subsequently, using this reagent, a pyrene-modified uridine is incorporated into RNA by using an automatic DNA synthesizer. |
