Wakunaga Seiyaku Kabushiki Kaisha

Kenichi Miyoshi

Toru Fuwa

Masanori Suzuki

Akio Yamane

Akira Hasegawa

Takanori Oka

Satoru Nakagami

Noriko Tsukumo

Tatsuro Kawasoe

Shinichiro Sumi

An intended nucleic acid is determined in a sample by a method in which at least one of the nucleic acid strands of the intended nucleic acid is subjected to hybridization with a primer and to chain-extension reaction over the primer to form a synthesized nucleic acid which is complementary to and in hybridization with the strand; a copy of the intended nucleic acid is obtained, which copy is (a) the nucleic acid of the double-stranded double produced by the chain-extension reaction, or (b) the synthesized nucleic acid freed from the double-stranded nucleic acid, or (c) a double stranded nucleic acid form from a pair of the synthesized nucleic acids complementary with each other; and it is determined whether the copy is present in the sample thereby to know whether the intended nucleic acid is in present in the sample.

Method for detecting target nucleic acid in specimen

A poly-labelled oligonucleotide having a plurality of labels, comprising labels carried on a polylysine and having the polylysine introduced on the extension from 5'- and/or 3'-end through phosphate group is useful as a probe for hybridization.

Poly-labelled oligonucleotide derivative

Biotin- or 2,4-dinitrophenyl-oligodeoxyribonucleotide derivative represented by the formula shown below and its preparation method is disclosed: ##STR1## wherein: R is ##STR2## m and n are each 0 or a natural number; R.sup.1 is a straight or branched hydrocarbon residue; and B is a base constituting the nucleotide (in which when a plural number of B exist, they may be either identical or different). This compound is useful for use as a nonradioactive affinity probe for nucleic acid or a primer.

Oligonucleotide derivatives and production thereof

An oligonucleotide derivative having an amino group protected with an eliminatable group bonded through a phosphate group and a spacer with an appropriate length to the 5'-end of an oligonucleotide protected suitably at the 3'-hydroxyl group and the base moiety of the nucleotide, and an immobilized oligonucleotide derivative having a Sepharose carrier bonded to the amino group in place of said protective group are disclosed. Methods for production of these derivatives are also disclosed.

A nucleotide of the formula [IV] shown below: ##STR1## wherein: X is hydrogen or --CO--(CH.sub.2).sub.n --CO--R.sup.3 ; each of m and n is an integer up to 20; R.sup.0 is a protective group of a phosphoric acid group, R.sup.1 is hydrogen or a protective group for a 5'-hydroxyl group, R.sup.2 is hydrogen or a protective group for an amino group, R.sup.3 is a carrier having an amino group as the functional group and bound at this functional group in this compound, Y is oxygen atom (--0--) or an imino group (--NH--). This nucleotide, when the X is --CO--(CH.sub.2).sub.n --CO--R.sup.3, has a carrier (R.sup.3) through a spacer (--CO--(CH.sub.2).sub.m --CO--) on extension from the amino group of deoxycytidine and therefore can be used as the starting material, namely the resin, for synthesizing a 3'-aminated oligonucleotide according to the solid phase method.

Nucleotide derivative

[21-Leucine] human urogastrone, hUG, comprising a polypeptide represented by the specified amino acid sequence has been found to be equivalent to the known hUG which has 21-Met and is disclosed together with its syntheses. Double-stranded polydeoxyribonucleotides comprising a structural gene for expressing the hUG, a recombinant plasmid comprising a structural gene for expressing the hUG, and E. coli transformed by the recombinant plasmid are also disclosed. Thanks to the substitution of Met in the known hUG by Leu, the peptide of hUG can now be produced by a conventional or typical method in which a genetically engineered precursor fused polypeptide is cleaved by cyanogen bromide at Met in the fused polypeptide.

[21-leucine] human urogastrone, corresponding gene, corresponding

Oligonucleotide derivatives

Biotin- or 2,4-dinitrophenyl-oligodeoxyribonucleotide derivatve represented by the formula shown below and its preparation method is disclosed: ##STR1## wherein: R is ##STR2## m and n are each 0 or a natural number; R.sup.1 is a straight or branched hydrocarbon residue; and B is a base constituting the nucleotide (in which when a plural number of B exist, they may be either identical or different). This compound is useful for use as a nonradioactive affinity probe for nucleic acid or a primer.

Growing community of inventors

Hiroshima, Japan

Kenichi Miyoshi