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通过重组过程从啮齿动物-人类基因组文库中获取人类DNA。

Retrieval of human DNA from rodent-human genomic libraries by a recombination process.

作者信息

Neve R L, Bruns G A, Dryja T P, Kurnit D M

出版信息

Gene. 1983 Sep;23(3):343-54. doi: 10.1016/0378-1119(83)90023-9.

Abstract

Human Alu repeat ("BLUR") sequences have been cloned into the mini-plasmid vector piVX. The resulting piBLUR clones have been used to rescue selectively, by recombination, bacteriophage carrying human DNA sequences from genomic libraries constructed using DNA from rodent-human somatic cell hybrids. piBLUR clones are able to retrieve human clones from such libraries because at least one Alu family repeat is present on most 15 to 20 kb fragments of human DNA and because of the relative species-specificity of the sequences comprising the Alu family. The rapid, selective plaque purification achieved results in the construction of a collection of recombinant phage carrying diverse human DNA inserts from a specific subset of the human karyotype. Subfragments of two recombinants rescued from a mouse-human somatic cell hybrid containing human chromosomes X, 10, 13, and 22 were mapped to human chromosomes X and 13, respectively, demonstrating the utility of this protocol for the isolation of human chromosome-specific DNA sequences from appropriate somatic cell hybrids.

摘要

人类Alu重复序列(“BLUR”)已被克隆到微型质粒载体piVX中。所得的piBLUR克隆已用于通过重组选择性拯救携带人类DNA序列的噬菌体,这些噬菌体来自使用啮齿动物-人类体细胞杂种的DNA构建的基因组文库。piBLUR克隆能够从此类文库中检索人类克隆,因为在大多数15至20 kb的人类DNA片段上至少存在一个Alu家族重复序列,并且由于构成Alu家族的序列具有相对物种特异性。所实现的快速、选择性噬菌斑纯化导致构建了一组携带来自人类核型特定子集的不同人类DNA插入片段的重组噬菌体。从含有人类染色体X、10、13和22的小鼠-人类体细胞杂种中拯救的两个重组体的亚片段分别被定位到人类染色体X和13上,证明了该方案对于从合适的体细胞杂种中分离人类染色体特异性DNA序列的实用性。

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