Receptor proteins specifically recognizing bacterial dna

Details Receptor proteins specifically recognizing bacterial dna Receptor proteins specifically recognizing bacterial dna

C - Chemistry, Metallurgy

12

N

C12N 15/12 (2006.01) A61K 38/00 (2006.01) A61K 45/00 (2006.01) A61P 31/04 (2006.01) A61P 35/00 (2006.01) A61P 37/08 (2006.01) C07K 14/705 (2006.01) C07K 16/28 (2006.01) C12N 5/10 (2006.01) C12P 21/02 (2006.01) C12P 21/08 (2006.01) C12Q 1/68 (2006.01) G01N 33/15 (2006.01) G01N 33/50 (2006.01) G01N 33/566 (2006.01) G01N 33/577 (2006.01)

Patent

CA 2385302

The present invention provides a receptor protein specifically recognizing bacterial DNA having an unmethylated CpG sequence, a genomic DNA encoding it, an experimental animal model useful for examining responsiveness of a host immune cell against a bacterial infectious disease. DNA encoding a receptor protein specifically recognizing bacterial DNA having an unmethylated CpG sequence is screened by BLAST search, a number of EST clones having high homology with various TLRs is screened, these clones are used as a probe to isolate a full-length cDNA from mouse macrophage cDNA library, and the sequence of bases of the cDNA is analyzed to confirm that it is TLR9 comprising a conserved regions such as LRR and TIR regions, and then a knockout mouse is produced to confirm that TLR9 is a receptor protein of oligonucleotides having an unmethylated CpG sequence of bacterial DNA.

La présente invention concerne, d'une part une protéine réceptrice reconnaissant spécifiquement un ADN bactérien disposant d'une séquence CpG non méthylée, un ADN génique codant cet ADN, et des modèles animaux convenant à l'étude des réponses immunitaire des immunocytes à des maladies infectieuses bactériennes. On commence par cribler, selon le procédé de recherche BLAST, un ADN codant une protéine réceptrice reconnaissant spécifiquement un ADN bactérien disposant d'une séquence CpG non méthylée. Puis on crible un certain nombre de clones EST hautement analogues à divers TLR. L'utilisation de ces clones comme sondes permet d'isoler un ADN complémentaire pleine longueur dans une échantillothèque d'ADN complémentaire de macrophages murins. Après avoir analysé la séquence de base de l'ADN complémentaire et confirmé qu'il s'agit de TLR9 ayant conservé des domaines tels que les LRR et TIR, une souris knock-out et construite. Il est ainsi confirmé que le TLR9 est une protéine réceptrice d'un oligonucléotide contenant la séquence CpG non méthylée d'un ADN bactérien.

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