Montoliu L, Schedl A, Kelsey G, Zentgraf H, Lichter P, Schütz G
Division of Molecular Biology of the Cell I, German Cancer Research Center, Heidelberg.
Reprod Fertil Dev. 1994;6(5):577-84. doi: 10.1071/rd9940577.
Several groups have recently reported the successful generation of transgenic mice harbouring yeast artificial chromosomes (YACs). Different methodological approaches have been shown to produce similar results, namely, the faithful expression of the transgenes carried on YAC DNA. In this paper, we compare the reported techniques for obtaining transgenic mice carrying YACs using a 250-kb YAC bearing the mouse tyrosinase gene. These methods include: microinjection of gel-purified YAC DNA into pronuclei of fertilized mouse oocytes, yeast spheroblast fusion with embryonic stem (ES) cells and lipofection of YAC DNA into ES cells. Taken together, these reports show that the delivery of large genomic regions covering a gene of interest (such as those cloned in YAC vectors) is feasible, and will ensure appropriate temporal and spatial expression of the transgene at a level comparable to that of the endogenous counterpart.
最近有几个研究小组报告了成功培育出携带酵母人工染色体（YAC）的转基因小鼠。已证明不同的方法能产生相似的结果，即YAC DNA上携带的转基因能够如实表达。在本文中，我们使用携带小鼠酪氨酸酶基因的250 kb YAC，比较了已报道的获得携带YAC的转基因小鼠的技术。这些方法包括：将凝胶纯化的YAC DNA显微注射到受精小鼠卵母细胞的原核中、酵母球状体与胚胎干细胞（ES细胞）融合以及将YAC DNA脂质转染到ES细胞中。综合这些报告表明，传递覆盖感兴趣基因的大片段基因组区域（如那些克隆在YAC载体中的区域）是可行的，并且将确保转基因在时间和空间上的表达与内源性对应物相当。
