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Substitution of the α-lactalbumin transcription unit by a CAT cDNA within a BAC clone silenced the locus in transgenic mice without affecting the physically linked Cyclin T1 gene

  • 1,
  • 1,
  • 1,
  • 2,
  • 2,
  • 1 and
  • 1Email author
Genetics Selection Evolution200335:239

https://doi.org/10.1186/1297-9686-35-2-239

  • Received: 12 December 2001
  • Accepted: 1 October 2002
  • Published:

Abstract

We recently reported that a goat bacterial artificial chromosome (BAC) clone conferred site-independent expression in transgenic mice of the two loci present within its insert, the ubiquitously expressed Cyclin T1 and the mammary specific β-lactalbumin (αlac) genes. To assess if this vector could target mammary-restricted expression of cDNA, the CAT ORF was introduced by homologous recombination in Escherichia coli in place of the αlac transcription unit. The insert of this modified BAC was injected into mice and three transgenic lines were derived. None of these lines expressed the CAT gene suggesting that the use of long genomic inserts is not sufficient to support the expression of intron-less transgenes. The physically linked goat Cyclin T1 locus was found to be active in all three lines. This observation reinforced the hypothesis that the two loci are localised in two separate chromatin domains.

Keywords

  • bacterial artificial chromosome
  • homologous recombination
  • intron
  • transgenic mice
  • chromatin domain

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Authors’ Affiliations

(1)
Laboratoire de génétique biochimique et de cytogénétique, Département de génétique animale, Institut national de la recherche agronomique, 78352 Jouy-en-Josas Cedex, France
(2)
Laboratoire de biologie du développement et de biotechnologie, Département de physiologie animale, Institut national de la recherche agronomique, 78352 Jouy-en-Josas Cedex, France

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