Scambler P J, Estivill X, Bell G, Farrall M, McLean C, Newman R, Little P F, Frederick P, Hawley K, Wainwright B J
Nucleic Acids Res. 1987 May 11;15(9):3639-52. doi: 10.1093/nar/15.9.3639.
Cosmid libraries have been constructed from DNA of somatic cell hybrid cell lines, each containing a fragment of human chromosome seven and including sequences closely linked to cystic fibrosis (CF). Cosmids containing human DNA as insert were isolated from the library. Three cosmids, when used as probes to total genomic DNA, detected polymorphic loci, each of which was shown to be in strong linkage disequilibrium with CF. Restriction endonuclease digestion of cosmid clones and use of a new, rapid method of chromosome walking based on competitive hybridisation of cosmid inserts has allowed identification of several groups of overlapping cosmids ("contigs") from the vicinity of CF.
黏粒文库是由体细胞杂交细胞系的DNA构建而成,每个细胞系都包含人类7号染色体的一个片段,并且包含与囊性纤维化(CF)紧密连锁的序列。从文库中分离出了以人类DNA为插入片段的黏粒。当将三个黏粒用作总基因组DNA的探针时,检测到了多态性位点,每个位点都显示与CF存在强连锁不平衡。对黏粒克隆进行限制性内切酶消化,并使用基于黏粒插入片段竞争性杂交的新的快速染色体步移方法,已从CF附近鉴定出几组重叠的黏粒(“重叠群”)。