Activator effect of coinjected enhancers on the muscle-specific expression of promoters in zebrafish embryos
Müller, F., Williams, D.W., Kobolák, J., Gauvry, L., Goldspink, G., Orbán, L. and Maclean, N. (1997) Activator effect of coinjected enhancers on the muscle-specific expression of promoters in zebrafish embryos. Molecular Reproduction and Development, 47 (4). pp. 404-412.
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The transient expression of reporter gene constructs in embryos provides a powerful tool to characterise cis-acting transcriptional elements of the genes involved in development.
In the present study, we have analysed the expression pattern of several muscle-specific and ubiquitous regulatory sequences in microinjected zebrafish embryos. By using a fast and reproducible coinjection strategy, the mosaic expression of lacZ reporter gene was monitored in wholemount embryos injected with sequences containing putative enhancer elements and a carp myosin heavy chain promoter/lacZ reporter construct. We have found that a 0.9-kb myosin heavy chain (MyHC) proximal promoter containing several putative myogenic regulatory factors (MRF) binding sites is sufficient to restrict lacZ expression to the skeletal muscle fibres of prim-6 stage zebrafish embryos. Expression of a rat-derived foetal myosin light chain enhancer (MyLC) and different fragments of a carp β-actin regulatory region together with the MyHC promoter were compared by accumulating the type, number and spatial distribution of β-galactosidase-expressing cells on an expression map. β-galactosidase activity increased similarly whether the MyLC enhancer was ligated to the promoter/reporter construct directly or when coinjected as a separate fragment whilst skeletal muscle specificity was retained. The coinjection of two different forms of the β-actin regulatory elements also showed a marked effect on the MyHC promoter activity.
|Publication Type:||Journal Article|
|Copyright:||© 1997 Wiley-Liss, Inc.|
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