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人类9号染色体黏粒文库的构建与鉴定

Generation and characterization of a human chromosome 9 cosmid library.

作者信息

Graw S L, Buckler A J, Britt D E, Jackson C L, Taruscio D, Baldini A, Ward D C, Housman D E

机构信息

Department of Biology, Massachusetts Institute of Technology, Cambridge 02139.

出版信息

Somat Cell Mol Genet. 1992 May;18(3):269-84. doi: 10.1007/BF01233863.

Abstract

A cosmid library has been constructed from the hamster-human hybrid cell line PK-87-9, which contains chromosome 9 as its sole known human component. Ten thousand colonies were produced, of which approximately 200, or 2%, contain human material. Fifty of these 200 were regionally mapped by an Alu-primed PCR product hybridization procedure. These cosmids were localized to all regions of chromosome 9, but were especially concentrated in the distal portion of 9q. The map location derived by the Alu-primed PCR product hybridization procedure was compared to the map location derived by fluorescent in situ hybridization. Assignment of chromosomal location by the two methods was correspondent in all but a few cases. The presumptive presence of HTF islands was investigated for 130 cosmids by digestion with the restriction enzyme NotI. Twenty percent of cosmids contained at least one NotI site. A number of simple sequence repeat polymorphisms identified from the cosmid set were characterized and will provide a link between the genetic and physical maps for this chromosome.

摘要

已从仓鼠 - 人杂交细胞系PK - 87 - 9构建了一个黏粒文库,该细胞系含有9号染色体作为其唯一已知的人类成分。产生了一万个菌落,其中约200个(即2%)含有人类物质。通过Alu引物PCR产物杂交程序对这200个中的50个进行了区域定位。这些黏粒定位于9号染色体的所有区域,但特别集中在9q的远端部分。将通过Alu引物PCR产物杂交程序得出的图谱位置与通过荧光原位杂交得出的图谱位置进行了比较。除少数情况外,两种方法对染色体位置的定位是一致的。通过用限制酶NotI消化,对130个黏粒的HTF岛的推测存在情况进行了研究。20%的黏粒含有至少一个NotI位点。从黏粒组中鉴定出的一些简单序列重复多态性进行了表征,它们将为该染色体的遗传图谱和物理图谱之间提供联系。

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