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通过杂交细胞DNA的分子克隆分离出人类7号染色体特有的多态性DNA片段。

Isolation of a polymorphic DNA segment unique to human chromosome 7 by molecular cloning of hybrid cell DNA.

作者信息

Humphries P, Barton D, McKay A M, Humphries M M, Carritt B

出版信息

Mol Gen Genet. 1983;190(1):143-9. doi: 10.1007/BF00330337.

Abstract

DNA isolated from a rodent-human hybrid cell line containing human chromosomes 3, 7, 9, 10, 14 and 22 was cloned in the plasmid vector pAT153. Recombinant plasmids containing inserts of human origin were identified by colony hybridization to 32P-labelled human DNA under conditions in which only repetitive sequences interact. Single- and low-copy sequences were liberated from these plasmids by restriction endonuclease digestion and used as hybridization probes against human DNA and DNA isolated from a panel of Chinese hamster-human hybrids. One single-copy probe was shown to react with a genomic sequence unique to human chromosome 7 and to recognize an apparent restriction fragment size polymorphism in human DNA.

摘要

从含有人类3号、7号、9号、10号、14号和22号染色体的啮齿动物-人类杂交细胞系中分离出的DNA,被克隆到质粒载体pAT153中。通过在仅重复序列相互作用的条件下与32P标记的人类DNA进行菌落杂交,鉴定出含有人类来源插入片段的重组质粒。通过限制性内切酶消化从这些质粒中释放出单拷贝和低拷贝序列,并将其用作针对人类DNA以及从一组中国仓鼠-人类杂交体中分离出的DNA的杂交探针。一个单拷贝探针被证明与人类7号染色体特有的基因组序列发生反应,并识别出人类DNA中明显的限制性片段大小多态性。

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