Genomic studies of gene expression: regulation of the Wilson disease gene

被引：6
作者：
Bochukova, EG
Jefferson, A
Francis, MJ
Monaco, AP [1 ]
机构：
[1] Univ Oxford, Wellcome Trust Ctr Human Genet, Oxford OX3 7BN, England
[2] Maynard Ctr, Amersham Biosci, Cardiff CF4 7YT, S Glam, Wales
来源：
GENOMICS | 2003年 / 81卷 / 06期
关键词：
bacterial artificial chromosomes; gene regulation; Wilson disease; genomic gene therapy;
D O I：
10.1016/S0888-7543(03)00093-4
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：
Bacterial artificial chromosomes (BACs) have many advantages over other large-insert cloning vectors and have been used for a variety of genetic Applications, including the final contigs of the human genome. We describe the utilization of a BAC construct to study gene regulation in a tissue culture-based system, using a 170-kb clone containing the entire Wilson disease (WND) locus as a model. A second BAC construct that lacked a putative negatively regulating promoter, sequence was created. A nonviral method of gene delivery was applied to transfect three human cell lines stably with each construct. Our results show correct WND gene expression from the recombinant locus and quantification revealed significantly increased expression from the clone lacking the negative regulator. Comparison with conventional methods confirms the reliability of the genomic approach for thorough examination of gene expression. This experimental system illustrates the potential of BAC clones in genomic gene expression studies, new gene therapy strategies, and validation of potential molecular targets for drug discovery. (C) 2003 Elsevier Science (USA). All rights reserved.
引用
收藏
页码：531 / 542
页数：12
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