Smoller D A, Petrov D, Hartl D L
Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110-1095.
Chromosoma. 1991 Sep;100(8):487-94. doi: 10.1007/BF00352199.
A multiple-hit bacteriophage P1 library containing DNA fragments from Drosophila melanogaster in the size range 75-100 kb was created and subjected to a preliminary evaluation for completeness, randomness, fidelity, and clone stability. This P1 library presently contains 3840 individual clones, or approximately two genome equivalents. The library was screened with a small set of unique-sequence test probes, and clones containing the sequences have been recovered. In situ hybridization with salivary gland chromosomes indicates that the clones originate from the site of the probe sequences in the genome, and filter hybridization of restriction digests suggests that the clones are not rearranged in comparison with the genomic sequences. Approximately 1.7% of the clones contain sequences that hybridize with ribosomal DNA. A small subset of these clones was tested for stability by examination of restriction fragments produced after repeated subculturing, and no evidence for instability was found. The P1 cloning system has general utility in molecular genetics and may provide an important intermediate level of resolution in physical mapping of the Drosophila genome.
构建了一个多重打击噬菌体P1文库,其中包含大小在75 - 100 kb范围内的黑腹果蝇DNA片段,并对其完整性、随机性、保真度和克隆稳定性进行了初步评估。这个P1文库目前包含3840个独立克隆,约相当于两个基因组当量。用一小套单拷贝序列测试探针筛选该文库,已回收了包含这些序列的克隆。与唾液腺染色体的原位杂交表明,这些克隆起源于基因组中探针序列的位点,限制性消化产物的滤膜杂交表明,与基因组序列相比,这些克隆没有重排。约1.7%的克隆包含与核糖体DNA杂交的序列。通过检查重复传代后产生的限制性片段,对这些克隆中的一小部分进行了稳定性测试,未发现不稳定的证据。P1克隆系统在分子遗传学中具有广泛的用途,可能在果蝇基因组的物理图谱绘制中提供重要的中等分辨率水平。