CONSTRUCTION AND CHARACTERIZATION OF A HUMAN-CHROMOSOME 2-SPECIFIC BAC LIBRARY

被引:20
作者
WANG, M
CHEN, XN
SHOUSE, S
MANSON, J
WU, QZ
LI, RM
WRESTLER, J
NOYA, D
SUN, ZG
KORENBERG, J
LAI, E
机构
[1] UNIV N CAROLINA,DEPT PHARMACOL,CHAPEL HILL,NC 27599
[2] UNIV CALIF LOS ANGELES,CEDARS SINAI MED CTR,AHMANSON DEPT PEDIAT,LOS ANGELES,CA 90048
关键词
D O I
10.1006/geno.1994.1662
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Fire have constructed a human chromosome 2-specific bacterial artificial chromosome (BAC) library using DNA from the somatic cell hybrid GM10826. The average size of the clones is about 63 kb. The coverage and distribution of the library were estimated by screening with known polymorphic genetic markers and fluorescence in situ hybridization (FISH). Twenty-one markers tested positive when DNA pools prepared from approximately one-sixth of the library were screened with 33 known markers. This is consistent with the theoretical calculation of 63% coverage at one genomic equivalent. This suggested that the coverage of the library is approximately 5-6x. FISH analysis with 54 BACs revealed single site hybridization to chromosome 2, and the clones were distributed randomly on the chromosome. We have also performed direct sequencing of the BAC insert ends to generate sequence-tagged sites suitable for mapping and chromosome walking. This is the first reported human chromosome 2-specific BAC Library and should provide a resource for physical mapping and disease searching for this chromosome. (C) 1994 Academic Press, Inc.
引用
收藏
页码:527 / 534
页数:8
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