Olsen A S, Georgescu A, Johnson S, Carrano A V
Human Genome Center, Lawrence Livermore National Laboratory, Livermore, California, 94551, USA.
Genomics. 1996 Jun 1;34(2):223-5. doi: 10.1006/geno.1996.0270.
We describe the assembly of a 1-Mb cosmid contig and restriction map spanning the candidate region for Finnish congenital nephrosis (NPHS1) in 19q13.1. The map was constructed from 16 smaller contigs assembled by fingerprinting, a BAC and a PAC clone, and 42 previously unmapped cosmids. In most cases, single-step cosmid walks were sufficient to join two previously assembled contigs, and all but one gap was filled from this cosmid contig library. The remaining gap of about 19 kb was spanned with a single BAC and a single PAC clone. EcoRI mapping of a dense set of overlapping clones validated the assembly of the map and indicated a length of 1040 kb for the contig. This high-resolution clone map provides an ideal resource for gene identification through cDNA selection, exon trapping, and DNA sequencing.
我们描述了一个跨越19q13.1上芬兰先天性肾病(NPHS1)候选区域的1兆碱基粘粒重叠群及限制性图谱的构建。该图谱由通过指纹分析组装的16个较小重叠群、一个BAC和一个PAC克隆以及42个先前未定位的粘粒构建而成。在大多数情况下,单步粘粒步移足以连接两个先前组装的重叠群,并且除了一个间隙外,所有间隙都从这个粘粒重叠群文库中填补。剩余约19 kb的间隙由一个单一的BAC和一个单一的PAC克隆跨越。一组密集的重叠克隆的EcoRI图谱验证了图谱的组装,并表明重叠群的长度为1040 kb。这个高分辨率克隆图谱为通过cDNA选择、外显子捕获和DNA测序进行基因鉴定提供了理想资源。