Deák P, Omar M M, Saunders R D, Pál M, Komonyi O, Szidonya J, Maróy P, Zhang Y, Ashburner M, Benos P, Savakis C, Siden-Kiamos I, Louis C, Bolshakov V N, Kafatos F C, Madueno E, Modolell J, Glover D M
Department of Anatomy and Physiology, University of Dundee, United Kingdom.
Genetics. 1997 Dec;147(4):1697-722. doi: 10.1093/genetics/147.4.1697.
We have established a collection of 2460 lethal or semi-lethal mutant lines using a procedure thought to insert single P elements into vital genes on the third chromosome of Drosophila melanogaster. More than 1200 randomly selected lines were examined by in situ hybridization and 90% found to contain single insertions at sites that mark 89% of all lettered subdivisions of the Bridges' map. A set of chromosomal deficiencies that collectively uncover approximately 25% of the euchromatin of chromosome 3 reveal lethal mutations in 468 lines corresponding to 145 complementation groups. We undertook a detailed analysis of the cytogenetic interval 86E-87F and identified 87 P-element-induced mutations falling into 38 complementation groups, 16 of which correspond to previously known genes. Twenty-one of these 38 complementation groups have at least one allele that has a P-element insertion at a position consistent with the cytogenetics of the locus. We have rescued P elements and flanking chromosomal sequences from the 86E-87F region in 35 lines with either lethal or genetically silent P insertions, and used these as probes to identify cosmids and P1 clones from the Drosophila genome projects. This has tied together the physical and genetic maps and has linked 44 previously identified cosmid contigs into seven "super-contigs" that span the interval. STS data for sequences flanking one side of the P-element insertions in 49 lines has identified insertions in the alphagamma element at 87C, two known transposable elements, and the open reading frames of seven putative single copy genes. These correspond to five known genes in this interval, and two genes identified by the homology of their predicted products to known proteins from other organisms.
我们利用一种被认为可将单个P因子插入黑腹果蝇第三条染色体上重要基因的方法,建立了一个包含2460个致死或半致死突变品系的文库。通过原位杂交对1200多个随机挑选的品系进行了检测,发现90%的品系在标记了布里奇斯图谱所有字母细分区域89%的位点上含有单个插入。一组共同覆盖约25%的3号染色体常染色质的染色体缺失,在468个品系中揭示了致死突变,这些品系对应于145个互补群。我们对细胞遗传学区间86E - 87F进行了详细分析,鉴定出87个P因子诱导的突变,分为38个互补群,其中16个对应于先前已知的基因。这38个互补群中的21个至少有一个等位基因,其P因子插入位置与该位点的细胞遗传学一致。我们从35个具有致死或遗传沉默P插入的品系中拯救了86E - 87F区域的P因子和侧翼染色体序列,并将其用作探针,从果蝇基因组计划中鉴定黏粒和P1克隆。这将物理图谱和遗传图谱联系在一起,并将44个先前鉴定的黏粒重叠群连接成跨越该区间的7个“超级重叠群”。49个品系中P因子插入一侧侧翼序列的STS数据,在87C处的alphagamma元件、两个已知的转座元件以及七个推定单拷贝基因的开放阅读框中鉴定出插入。这些对应于该区间的五个已知基因,以及两个通过其预测产物与其他生物体已知蛋白质的同源性鉴定的基因。