GERM LINE TRANSMISSION OF A YEAST ARTIFICIAL CHROMOSOME SPANNING THE MURINE ALPHA(1)(I) COLLAGEN LOCUS
GERM LINE TRANSMISSION OF A YEAST ARTIFICIAL CHROMOSOME SPANNING THE MURINE ALPHA(1)(I) COLLAGEN LOCUS
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DOI：
10.1126/science.8096090
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发表时间：
1993-03-26
期刊：
影响因子：
56.9
通讯作者：
JAENISCH, R
中科院分区：
文献类型：
--
作者：
STRAUSS, WM;DAUSMAN, J;JAENISCH, R
利用转基因技术实现突变表型的分子互补是一种潜在的重要基因鉴定工具。开发了一种技术，可以将物理上完整的酵母人工染色体（YAC）转移到小鼠的种系中。将含有小鼠基因Col1a1的纯化的150千碱基YAC通过脂肪转染有效地导入胚胎干细胞。嵌合创始人小鼠来源于两个转染的胚胎干细胞克隆。这些嵌合体通过种系传递了全长的转基因，产生了两个转基因小鼠品系。在间期细胞核中以新生转录本的形式显示转基因表达，并通过核糖核酸酶保护分析进行定量分析。两项实验都表明，转基因的表达水平与内源性胶原蛋白基因相当。
Molecular complementation of mutant phenotypes by transgenic technology is a potentially important tool for gene identification. A technology was developed that allows the transfer of a physically intact yeast artificial chromosome (YAC) into the germ line of the mouse. A purified 150-kilobase YAC encompassing the murine gene Col1a1 was efficiently introduced into embryonic stem (ES) cells via lipofection. Chimeric founder mice were derived from two transfected ES cell clones. These chimeras transmitted the full length transgene through the germ line, generating two transgenic mouse strains. Transgene expression was visualized as nascent transcripts in interphase nuclei and quantitated by ribonuclease protection analysis. Both assays indicated that the transgene was expressed at levels comparable to the endogenous collagen gene.