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来自显微切割的人类染色体区域15q11 - q13的黏粒克隆

Cosmid clones from microdissected human chromosomal region 15q11-q13.

作者信息

Tohma T, Tamura T, Ohta T, Soejima H, Kubota T, Jinno Y, Tsukamoto K, Nakamura Y, Naritomi K, Niikawa N

机构信息

Department of Pediatrics, University of the Ryukyus School of Medicine, Okinawa, Japan.

出版信息

Jpn J Hum Genet. 1993 Sep;38(3):267-75. doi: 10.1007/BF01874137.

Abstract

A human chromosomal region, 15q11-q13, was microdissected, its DNA was amplified with the primer-linker PCR method, and the PCR products were cloned into a plasmid vector to construct a microclone library. Of 193 microclones analyzed with Southern blot hybridization on hybrid cell panels, 26 (13.5%) were either single-copy (unique) or low-repetitive fragments. By screening of a cosmid library of human genomic DNA using the 26 microclones as probes, 47 positive cosmids were obtained and underwent regional mapping with chromosome fluorescence in situ hybridization (FISH). Sixteen cosmids gave FISH signals at 15p-cen, 5 at 15q11-q13, 6 at 15q22-q26, 3 at other chromosomes, and 17 no signal. These 27 cosmids mapped to chromosome 15 are useful additions to the inventory of DNA markers of this chromosome including the much interested Prader-Willi/Angelman syndrome region.

摘要

对人类染色体区域15q11 - q13进行显微切割,采用引物连接PCR方法扩增其DNA,并将PCR产物克隆到质粒载体中构建微克隆文库。在用杂交细胞板进行Southern印迹杂交分析的193个微克隆中,26个(13.5%)为单拷贝(独特)或低重复片段。以这26个微克隆为探针筛选人类基因组DNA的黏粒文库,获得47个阳性黏粒,并通过染色体荧光原位杂交(FISH)进行区域定位。16个黏粒在15p - cen处产生FISH信号,5个在15q11 - q13处,6个在15q22 - q26处,3个在其他染色体上,17个无信号。这27个定位到15号染色体的黏粒是该染色体DNA标记库的有用补充,包括备受关注的普拉德 - 威利/安吉尔曼综合征区域。

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