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Predicate | Object |
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
4
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pubmed:dateCreated |
1997-8-18
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pubmed:abstractText |
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 beta-actin regulatory region together with the MyHC promoter were compared by accumulating the type, number and spatial distribution of beta-galactosidase-expressing cells on an expression map. beta-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 beta-actin regulatory elements also showed a marked effect on the MyHC promoter activity. The coinjection of putative enhancers with minimal promoter constructs and subsequent analysis of the transient expression pattern in the developing embryos provides a rapid and simple technique to identify cis acting activator elements of genes expressed in the vertebrate embryo.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Aug
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pubmed:issn |
1040-452X
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:volume |
47
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
404-12
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pubmed:dateRevised |
2008-11-21
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pubmed:meshHeading |
pubmed-meshheading:9211424-Actins,
pubmed-meshheading:9211424-Animals,
pubmed-meshheading:9211424-Embryo, Nonmammalian,
pubmed-meshheading:9211424-Enhancer Elements, Genetic,
pubmed-meshheading:9211424-Gene Expression Regulation, Developmental,
pubmed-meshheading:9211424-Genes, Reporter,
pubmed-meshheading:9211424-Histocytochemistry,
pubmed-meshheading:9211424-Lac Operon,
pubmed-meshheading:9211424-Microinjections,
pubmed-meshheading:9211424-Muscle, Skeletal,
pubmed-meshheading:9211424-Myosin Heavy Chains,
pubmed-meshheading:9211424-Myosin Light Chains,
pubmed-meshheading:9211424-Promoter Regions, Genetic,
pubmed-meshheading:9211424-Rats,
pubmed-meshheading:9211424-Zebrafish,
pubmed-meshheading:9211424-beta-Galactosidase
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pubmed:year |
1997
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pubmed:articleTitle |
Activator effect of coinjected enhancers on the muscle-specific expression of promoters in zebrafish embryos.
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pubmed:affiliation |
Institute for Molecular Genetics, Agricultural Biotechnology Center, Gödöllö, Hungary.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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