Expression of human smooth muscle calponin in transgenic mice revealed with a bacterial artificial chromosome

被引:11
作者
Miano, JM
Kitchen, CM
Chen, JY
Maltby, KM
Kelly, LA
Weiler, H
Krahe, R
Ashworth, LK
Garcia, E
机构
[1] Univ Rochester, Med Ctr, Cardiovasc Res Ctr, Rochester, NY 14642 USA
[2] Med Coll Wisconsin, Dept Physiol, Milwaukee, WI 53226 USA
[3] Blood Ctr SE Wisconsin Inc, Milwaukee, WI 53201 USA
[4] Ohio State Univ, Div Human Canc Genet, Columbus, OH 43210 USA
[5] Lawrence Livermore Natl Lab, Ctr Human Genome, Livermore, CA 94550 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY | 2002年 / 282卷 / 05期
关键词
promoter; development; genome;
D O I
10.1152/ajpheart.00875.2001
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Defining regulatory elements governing cell-restricted gene expression can be difficult because cis-elements may reside tens of kilobases away from start site(s) of transcription. Artificial chromosomes, which harbor hundreds of kilobases of genomic DNA, preserve a large sequence landscape containing most, if not all, regulatory elements controlling the expression of a particular gene. Here, we report on the use of a bacterial artificial chromosome (BAC) to begin understanding the in vivo regulation of smooth muscle calponin (SM-Calp). Long and accurate polymerase chain reaction, sequencing, and in silico analyses facilitated the complete sequence annotation of a BAC harboring human SM-Calp (hSM-Calp). RNase protection, in situ hybridization, Western blotting, and immunohistochemistry assays showed the BAC clone faithfully expressed hSM-Calp in both cultured cells and transgenic mice. Moreover, expression of hSM-Calp mirrored that of endogenous mouse SM-Calp suggesting that all cis-regulatory elements governing hSM-Calp expression in vivo were contained within the BAC. These BAC mice represent a new model system in which to systematically assess regulatory elements governing SM-Calp transcription in vivo.
引用
收藏
页码:H1793 / H1803
页数:11
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