Methods for obtaining gene tags

Details Methods for obtaining gene tags Methods for obtaining gene tags

C - Chemistry, Metallurgy

12

N

C12N 15/09 (2006.01) C07K 14/47 (2006.01) C07K 16/18 (2006.01) C12N 1/15 (2006.01) C12N 1/19 (2006.01) C12N 1/21 (2006.01) C12N 5/10 (2006.01) C12N 15/12 (2006.01) C12P 21/02 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2547885

It is intended to provide a method of forming the 5~-end base sequence of an mRNA as a tag. Namely, a method involving the step of synthesizing a cDNA by using, as a template, an mRNA having a IIs linker containing the recognition sequence of a IIs type restriction enzyme in its CAP structure attached thereto. By treating this cDNA with the IIs type restriction enzyme, a tag comprising the 5~-end base sequence of the mRNA is formed. Thus, a tag can be formed from any mRNA without depending on base sequences. Based on the base sequence data of the above tag, it is possible to provide a method of identifying a transcription initiation point or a primer for synthesizing a full-length cDNA.

L'invention porte sur un procédé qui permet de transformer la séquence de base de l'extrémité 5' d'un ARMm en étiquette. L'invention concerne, en particulier, un procédé selon lequel on synthétise un ADNc en utilisant comme gabarit un ARNm comprenant un lieur IIs contenant la séquence de reconnaissance d'une enzyme de restriction de type IIs dans la structure CAP qui lui est attachée.En traitant l'ADNc avec l'enzyme de restriction de type IIs, on forme une étiquette comprenant la séquence de base de l'extrémité 5' de l'ARNm. De cette manière, on peut former un étiquette à partir de n'importe quel ARNm, indépendamment des séquences de base. Le procédé de l'invention permet, sur la base des données de la séquence de base de l'étiquette précitée, d'identifier un point de début de transcription ou une amorce pour la synthèse d'un ADNc de pleine longueur.

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