Sanford J, Kim B W, Deaven L L, Jones C, Higgins M J, Nowak N J, Shows T B
Department of Human Genetics, Roswell Park Cancer Institute, New York State Department of Health, Buffalo 14263.
Genomics. 1993 Mar;15(3):653-8. doi: 10.1006/geno.1993.1120.
A NotI end clone library has been constructed from a human-hamster hybrid cell line containing only human chromosome 11. Fifty-one NotI clones were chosen to characterize the library. The majority of NotI clones hybridize to small 15- to 200-kb fragments and have proven to be valuable for chromosome 11 physical mapping by detecting fragments not previously recognized by random probes. These NotI end clones have been used to isolate corresponding NotI linking cosmids which were then used to identify adjacent NotI fragments on pulsed-field gels. The clones were mapped using fluorescence in situ hybridization and a somatic cell hybrid panel. Although these clones were localized over the entirety of chromosome 11, a nonrandom distribution was observed. Northern blot analysis indicated that 57% (17/30) of the NotI clones examined detected poly(A)+ transcripts in HeLa cell RNA.
已从仅包含人类11号染色体的人-仓鼠杂交细胞系构建了一个NotI末端克隆文库。选择了51个NotI克隆来表征该文库。大多数NotI克隆与15至200 kb的小片段杂交，并已证明通过检测随机探针以前未识别的片段，对11号染色体物理图谱绘制很有价值。这些NotI末端克隆已用于分离相应的NotI连接黏粒，然后用于在脉冲场凝胶上鉴定相邻的NotI片段。使用荧光原位杂交和体细胞杂交板对克隆进行定位。尽管这些克隆定位在11号染色体的整个区域，但观察到非随机分布。Northern印迹分析表明，在所检测的NotI克隆中，57%（17/30）在HeLa细胞RNA中检测到多聚腺苷酸加尾转录本。
