Targeted gene disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein.
Targeted gene disruption of the endogenous c-abl locus by homologous recombination with DNA encoding a selectable fusion protein.
复制标题
通过与编码选择性融合蛋白的 DNA 同源重组，对内源性 c-abl 基因座进行靶向基因破坏。
DOI：
10.1073/pnas.87.8.3210
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发表时间：
1990
影响因子：
11.1
通讯作者：
Goff,SP
中科院分区：
文献类型：
--
作者：
Schwartzberg,PL;Robertson,EJ;Goff,SP
我们已经引入了一个替代突变的c-abl基因座的小鼠胚胎干细胞之间的同源重组外源添加的DNA和内源性基因。最初生成的模型构建体由插入完整Abelson鼠白血病病毒基因组的v-abl基因中的无启动子的可选择的新霉素抗性标记组成，设计为表达为融合蛋白或通过翻译重启。对这些病毒基因组进行的v-abl和neo标志物传递的测试显示，在蛋白融合构建体中共表达更稳定。将neo融合物从该v-abl构建体亚克隆到无启动子的c-abl片段中，并将所得DNA用于转化胚胎干细胞。基因组DNA的直接筛选表明，高比例的耐药克隆产生的同源重组到内源性c-abl基因座。
We have introduced a substitution mutation into the c-abl locus of murine embryonic stem cells by homologous recombination between exogenously added DNA and the endogenous gene. Model constructs were initially generated that consisted of a promoterless selectable neomycin resistance marker inserted into the v-abl gene of the complete Abelson murine leukemia virus genome, designed to be expressed either as a fusion protein or by translational restart. Tests of these viral genomes for transmission of v-abl and neo markers showed more stable coexpression in a protein fusion construct. The neo fusion was subcloned from this v-abl construct into a promoterless c-abl fragment, and the resulting DNA was used to transform embryonic stem cells. Direct screening of genomic DNAs showed that a high proportion of drug-resistant clones arose from homologous recombination into the endogenous c-abl locus.