Eiken Kagaku Kabushiki Kaisha

Eiken Kagaku Kabushiki Kaisya

Kentaro Nagamine

Tsugunori Notomi

Hidetoshi Kanda

Toshihiro Yonekawa

Yasuyoshi Mori

Nobuyoshi Shimizu

The present invention provides a nucleic acid synthesis method which involves the step of incubating a double-stranded nucleic acid template under conditions that ensure a complementary strand synthesis reaction using a primer as an origin. This method involves the step of placing a region, to which a primer capable of isothermally amplifying the template nucleic acid will anneal, in a condition that ensures base pairing, using an arbitrary primer. The arbitrary primer initiates the complementary strand synthesis reaction, using the double-stranded nucleic acid as a template and DNA polymerases catalyzing the complementary strand synthesis reaction which comprises the destabilization of the double-stranded nucleic acid and strand displacement, thereby providing a region that can undergo base pairing.

Method of amplifying nucleic acid by using double-stranded nucleic acid as template

The present invention realized isothermal and rapid polynucleotide synthesis by using as templates polynucleotides having a structure capable of forming loops, and combining a plurality of primers capable of providing a starting point for complementary strand synthesis to such loops. If the LAMP method is applied, all reactions can be carried out isothermally and rapidly since the template polynucleotides themselves can also be synthesized by an isothermal reaction.

Kit for synthesizing polynucleotides

Method for synthesizing polynucleotides

A method for detecting the occurrence of nucleic acid syntheses using an enzyme through the use of a generated insoluble substance as an indicator.

Method for detecting reaction product of nucleic acid synthesis

The method of the present invention is based on a method wherein a nucleic acid is synthesized utilizing the hybridized 3'-end, which is synthesized by complementary strand synthesis, on a specific region of a target nucleotide sequence existing as the nucleotide sequence of the same strand as the origin for the next round of complementary strand synthesis. The hybridization to a specific region, which region is searched for mutations and polymorphisms, is repeatedly carried out according to the present invention. Thus, mutations and polymorphisms in a target nucleotide sequence are exactly copied to the reaction products.

Method for detecting mutations and/or polymorphisms

Method for detecting product of nucleic acid synthesizing reaction

Method of detecting variation or polymorphism

A gene for a novel transmembrane protein was isolated and its nucleotide sequence was determined. A protein encoded by this gene (TRPC7) has 7 transmembrane regions, and because there is homology with TRP (transient receptor potential), this protein is considered to have calcium channel functions. Further, since a mutation in the TRPC7 gene is frequently observed in patients with manic-depressives insanity, TRPC7 is considered to be a pathogenic gene for bipolar affective disorder.

Growing community of inventors

Tochigi, Japan

Kentaro Nagamine