C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 7/01 (2006.01) A61K 35/76 (2006.01) A61K 48/00 (2006.01) A61P 35/00 (2006.01) C12N 15/869 (2006.01)
Patent
CA 2561691
It is intended to provide a method of quickly and surely preparing a recombinant herpes simplex virus (HSV) whereby a target protein can be expressed in cancer cells. This method of constructing a recombinant HSV comprises: the first step of inserting a BAC plasmid, which has a loxP site and an FRT site and has at least one expression cassette of a marker gene inserted between the loxP site and the FRT site into the herpes simplex virus (HSV) genome; the second step of constructing a shuttle vector having at least one expression cassette of a gene encoding the target protein, at least one marker gene and a loxP site and an FRT site individually inserted thereinto and inserting the shuttle vector into the loxP site of the HSV genome with the use of Cre recombinase; and the third step of coinfecting a host with the HSV genome and a vector capable of expressing Flp recombinase, excising the area between the FRT sites on the genome, and thus producing the target transgenic HSV.
La présente invention vise ·fournir un procéd·de prration rapide et sûre d'un virus de l'herpès simplex (H) recombinant selon lequel une protéine cible peut être exprimée s des cellules cancéreuses. Ce procéd·de construction d.apprxeq.un HSV recombinant comprend: d'abord l'insertion d'un plasmide BAC, lequel a un site loxP et un site FRT et a au moins une cassette d'expression d'un gt un site loxP et un site FRT insérés séparément dans celui-ci et l'insertion ecteur navette dans le site loxP du génome du HSV ·l'ide de Cre-recombinase; et enfin l'injection conjointe d'un hôte avec le génome du HSV et un vecteur capable d'exprer la Flp-recombinase, l'excision la zone entre les sites FRT présents sur le génome entraû ant ainsi la production du HSV trgénique cible.
Fukuhara Hiroshi
Todo Tomoki
Bereskin & Parr Llp/s.e.n.c.r.l.,s.r.l.
Todo Tomoki
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