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

Joseph M. Miano, Chad M. Kitchen, Jiyuan Chen, Kathleen M. Maltby, Louise A. Kelly, Hartmut Weiler, Ralf Krahe, Linda K. Ashworth, Emilio Garcia

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)H1793-H1803
JournalAmerican Journal of Physiology - Heart and Circulatory Physiology
Volume282
Issue number5 51-5
DOIs
StatePublished - 2002
Externally publishedYes

Keywords

  • Development
  • Genome
  • Promoter

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

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