Isolation of single-copy human genes from a library of yeast artificial chromosome clones.
Isolation of single-copy human genes from a library of yeast artificial chromosome clones.
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从酵母人工染色体克隆文库中分离单拷贝人类基因。
DOI：
10.1126/science.2544027
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Olson,MV
中科院分区：
文献类型：
--
作者：
Brownstein,BH;Silverman,GA;Little,RD;Burke,DT;Korsmeyer,SJ;Schlessinger,D;Olson,MV
最近发展起来的一种克隆系统，基于在酵母（酿酒酵母）中作为线性人工染色体的大分子DNA的繁殖，提供了一种以数百个碱基对为片段克隆整个人类基因组的潜在方法。该系统的大多数应用将需要能够从酵母人工染色体克隆文库中恢复特定序列，并在酵母中不加改变地繁殖这些序列。两个单拷贝基因现在已经从一个酵母人工染色体克隆文库中克隆出来，该文库是由人类全部DNA制备的。获得了多个独立的编码因子IX和纤溶酶原激活物抑制剂2型的基因。这些克隆的大小从60到650千碱基不等，在酵母中长时间繁殖是稳定的，并且似乎包含了人类DNA的忠实复制品。
A recently developed cloning system based on the propagation of large DNA molecules as linear, artificial chromosomes in the yeastSaccharomyces cerevisiaeprovides a potential method of cloning the entire human genome in segments of several hundred kilobase pairs. Most applications 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.