Huxley C, Gnirke A
Department of Genetics, Washington University Medical School, St. Louis, MO 63110.
Bioessays. 1991 Oct;13(10):545-50. doi: 10.1002/bies.950131009.
Human DNA can be cloned as yeast artificial chromosomes (YACs), each of which contains several hundred kilobases of human DNA. This DNA can be manipulated in the yeast host using homologous recombination and yeast selectable markers. In relatively few steps it is possible to make virtually any change in the cloned human DNA from single base pair changes to deletions and insertions. In order to study the function of the cloned DNA and the effects of the changes made in the yeast, the human DNA must be transferred back into mammalian cells. Recent experiments indicate that large genes can be transferred from the yeast host to mammalian cells in tissue culture and that the genes are transferred intact and are expressed. Using the same methods it may soon be possible to transfer YAC DNA into the mouse germ line so that the expression and function of genes cloned in YACs can be studied in developing and adult mammalian animals.
人类DNA可以被克隆为酵母人工染色体（YAC），每个YAC都包含几百千碱基的人类DNA。这种DNA可以在酵母宿主中利用同源重组和酵母选择标记进行操作。通过相对较少的步骤，就有可能对克隆的人类DNA进行几乎任何改变，从单碱基对改变到缺失和插入。为了研究克隆DNA的功能以及在酵母中所做改变的影响，必须将人类DNA再转移回哺乳动物细胞中。最近的实验表明，大基因可以从酵母宿主转移到组织培养中的哺乳动物细胞中，并且这些基因能够完整地转移并表达。使用同样的方法，不久之后或许就能够将YAC DNA转移到小鼠生殖系中，从而在发育中的和成年的哺乳动物体内研究克隆在YAC中的基因的表达和功能。
