Brownstein B H, Silverman G A, Little R D, Burke D T, Korsmeyer S J, Schlessinger D, Olson M V
Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110.
Science. 1989 Jun 16;244(4910):1348-51. doi: 10.1126/science.2544027.
A recently developed cloning system based on the propagation of large DNA molecules as linear, artificial chromosomes in the yeast Saccharomyces cerevisiae provides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. Most application of this system will require the ability to recover specific sequences from libraries of yeast artificial chromosome clones and to propagate these sequences in yeast without alterations. Two single-copy genes have now been cloned from a library of yeast artificial chromosome clones that was prepared from total human DNA. Multiple, independent isolates were obtained of the genes encoding factor IX and plasminogen activator inhibitor type 2. The clones, which ranged in size from 60 to 650 kilobases, were stable on prolonged propagation in yeast and appear to contain faithful replicas of human DNA.
最近开发的一种克隆系统，该系统基于将大型DNA分子作为线性人工染色体在酿酒酵母中进行繁殖，它提供了一种潜在的方法，可将整个人类基因组克隆成几百千碱基对的片段。该系统的大多数应用将需要具备从酵母人工染色体克隆文库中回收特定序列，并在酵母中无改变地繁殖这些序列的能力。现在已经从一个由总人类DNA制备的酵母人工染色体克隆文库中克隆出了两个单拷贝基因。获得了编码因子IX和纤溶酶原激活物抑制剂2型的基因的多个独立分离株。这些克隆大小在60至650千碱基之间，在酵母中长时间繁殖时是稳定的，并且似乎包含人类DNA的忠实复制品。
