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Nucleic Acids Research, 1988, Vol. 16, No. 4 1251-1271
© 1988


Articles

Functional activity of the two promoters of the myosin alkali light chain gene in primary muscle cell cultures: comparison with other muscle gene promoters and other culture systems

Philippe Daubas*, André Klarsfeld1, Ian Garner, Christian Pinset, Roger Cox and Margaret Buckingham

Unité de Génétique Moléculaire du Développement, Département de Biologie Moléculaire, Unité de Neurobiologie Moléculaire and Unité associée au CNRS UA041149, Intéractions Moléculaires et Cellulaires 1Département des Biotechnologies, Institut Pasteur 25, rue du docteur Roux, 75724 Paris Cedex, France

*To whom correspondence should be addressed

Received January 7, 1988. Accepted January 28, 1988.

Proximal upstream flanking sequences of the mouse myosin alkali light chain gene encoding MLC1F and MLC3F, the mouse {alpha}-cardiac actin gene and the chicken gene for the {alpha}-subunit of the acetylcholine receptor were linked to the bacterial chioramphenicol acetyl transferase (CAT) gene and transfected into primary cultures derived from mouse skeletal muscle or into myogenic cell lines. We demonstrate that the mouse MLC1F/MLC3F gene has two functional promoters. In primary muscle cultures, a 1200 bp sequence flanking exon 1 (MLC1F) and a 438 bp sequence flanking exon 2 (MLC3F) direct CAT activity in myotubes, but not in myoblasts or in non myogenic 3T6 and CV1 cells. Developmentally regulated expression is also seen with the {alpha}-cardiac actin (320 bp) and acetylcholine receptor {alpha}-subunit (850 bp) upstream sequences in the primary culture system. Transfection experiments with myogenic cell lines show different results with a given promoter construct, reflecting possible differences in the levels of regulatory factors between lines. Different muscle gene promoters behave differently in a given cell line, suggesting different regulatory factor requirements between these promoters.


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