Phage bonded to nuclear location signal

Details Phage bonded to nuclear location signal Phage bonded to nuclear location signal

C - Chemistry, Metallurgy

12

N

C12N 15/86 (2006.01) A61K 31/70 (2006.01) A61K 48/00 (2006.01) C07K 14/01 (2006.01) C07K 14/47 (2006.01) C07K 19/00 (2006.01) C12N 7/01 (2006.01) C12N 15/12 (2006.01) C12N 15/34 (2006.01) C12N 15/62 (2006.01) C12N 15/79 (2006.01) C12N 15/85 (2006.01) C12N 15/87 (2006.01)

Patent

CA 2262529

A .lambda. phage having a nuclear location signal imparted thereto has been obtained by constructing a vector capable of expressing a fused protein composed of a gpD protein constituting the head of a .lambda. phage with a nuclear location signal sequence, transforming Escherichia coli by this vector and, further, propagating a mutant .lambda. phage which cannot express the gpD protein in E. coli in this transformant. It has been confirmed that the obtained .lambda. phage is capable of packaging .lambda. phage DNAs of 80 % and 100 % in genome size. After further confirming that the nuclear location signal is exposed to the outside from the head of this phage, this phage has been micro-injected into cells to analyze its location activity toward the nuclei. Thus it has been clarified that this phage has a location activity toward nuclei.

Pour obtenir un phage .lambda. comportant un signal de localisation nucléaire, on a construit un vecteur capable d'exprimer une protéine de fusion constituée d'une protéine gpD formant la tête d'un phage .lambda. et d'une séquence du signal de localisation nucléaire, on a transformé Escherichia coli par ledit vecteur, puis on a propagé dans ce transformant un phage .lambda. mutant incapable d'exprimer ladite protéine gpD chez E. coli. Il a été confirmé que le phage .lambda. obtenu est capable d'emballer les ADN des phages .lambda. ayant un génome d'une taille égale à 80 et 100 %. Après avoir encore confirmé que le signal de localisation nucléaire est exposé à partir de la tête dudit phage, on a injecté ce dernier dans des cellules afin d'analyser son activité de localisation envers les noyaux. Ceci a permis de prouver qu'il possède effectivement une telle activité.

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