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    • 4. 发明申请
    • Method For Constructing Recombinant Herpes Simplex Virus
    • 构建重组单纯疱疹病毒的方法
    • US20070196336A1
    • 2007-08-23
    • US10594962
    • 2005-03-31
    • Tomoki TodoHiroshi Fukuhara
    • Tomoki TodoHiroshi Fukuhara
    • A61K48/00C12N15/86C12N7/00
    • C12N15/86A61K38/1774A61K38/20A61K38/208C12N7/00C12N2710/16632C12N2710/16643C12N2800/202C12N2800/30
    • A method is provided for rapidly and reliably constructing recombinant herpes simplex virus (HSV) capable of expressing a target protein in cancer cells. The method for constructing recombinant HSV as claimed in the present invention comprises a first step of inserting into a herpes simplex virus (HSV) genome, a BAC plasmid, which has a loxP site and an FRT site and into which has been inserted at least one type of marker gene expression cassette between the loxP site and the FRP site, a second step of constructing a shuttle vector into which has been respectively inserted at least one type of expression cassette of a gene encoding a target protein, at least one type of marker gene, a loxP site and an FRP site, and inserting the shuttle vector into the loxP site of the HSV genome using Cre recombinase, and a third step of co-infecting a host with the HSV genome and a vector capable of expressing Flp recombinase, and excising the region between the FRT sites in the genome to produce a target recombinant HSV.
    • 提供了一种快速可靠地构建能够在癌细胞中表达靶蛋白的重组单纯疱疹病毒(HSV)的方法。 用于构建本发明所要求的重组HSV的方法包括将第一步骤插入到单纯疱疹病毒(HSV)基因组中,BAC质粒具有loxP位点和FRT位点,其中插入有至少一个 类型的loxP位点和FRP位点之间的标记基因表达盒,构建穿梭载体的第二步,其中分别插入至少一种编码靶蛋白的基因的表达盒,至少一种类型的标记 基因,loxP位点和FRP位点,并使用Cre重组酶将穿梭载体插入HSV基因组的loxP位点,以及第三步,共同感染宿主与HSV基因组和能够表达Flp重组酶的载体, 并切除基因组中的FRT位点之间的区域以产生靶重组HSV。