Garza D, Ajioka J W, Burke D T, Hartl D L
Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110-1095.
Science. 1989 Nov 3;246(4930):641-6. doi: 10.1126/science.2510296.
The ability to clone large fragments of DNA in yeast artificial chromosomes (YAC's) has created the possibility of obtaining global physical maps of complex genomes. For this application to be feasible, most sequences in complex genomes must be able to be cloned in YAC's, and most clones must be genetically stable and colinear with the genomic sequences from which they originated (that is, not liable to undergo rearrangement). These requirements have been met with a YAC library containing DNA fragments from Drosophila melanogaster ranging in size up to several hundred kilobase pairs. Preliminary characterization of the Drosophila YAC library was carried out by in situ hybridization of random clones and analysis of clones containing known sequences. The results suggest that most euchromatic sequences can be cloned. The library also contains clones in which the inserted DNA is derived from the centromeric heterochromatin. The locations of 58 clones collectively representing about 8 percent of the euchromatic genome are presented.
利用酵母人工染色体（YAC）克隆大片段DNA的能力为获得复杂基因组的整体物理图谱创造了可能性。要使这种应用可行，复杂基因组中的大多数序列必须能够在YAC中克隆，并且大多数克隆必须在遗传上稳定，并且与它们所源自的基因组序列共线（即不易发生重排）。一个包含来自黑腹果蝇的大小达几百千碱基对的DNA片段的YAC文库满足了这些要求。通过对随机克隆进行原位杂交以及对包含已知序列的克隆进行分析，对果蝇YAC文库进行了初步表征。结果表明，大多数常染色质序列都可以被克隆。该文库还包含一些克隆，其中插入的DNA来自着丝粒异染色质。本文展示了58个克隆的位置，这些克隆共同代表了约8%的常染色质基因组。
