Linked Life Data

9441668

Source:http://linkedlifedata.com/resource/pubmed/id/9441668

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
1998-2-13
pubmed:abstractText
Despite a number of reports on transgenic zebrafish, there have been no reports on transgenic zebrafish in which the gene is under the control of a promoter of zebrafish origin. Neither have there been reports on transgenic zebrafish in which the gene is under the control of a tissue-specific promoter/enhancer. To investigate whether it is possible to generate transgenic zebrafish which reliably express a reporter gene in specific tissues, we have isolated a zebrafish muscle-specific actin (alpha-actin) promoter and generated transgenic zebrafish in which the green fluorescent protein (GFP) reporter gene was driven by this promoter. In total, 41 GFP-expressing transgenic lines were generated with a frequency of as high as 21% (41 of 194), and GFP was specifically expressed throughout muscle cells in virtually all of the lines (40 of 41). Nonexpressing transgenic lines were rare. This demonstrates that a tissue-specific promoter can reliably drive reporter gene expression in transgenic zebrafish in a manner identical to the control of the endogeneous expression of the gene. Levels of GFP expression varied greatly from line to line; i.e., fluorescence was very weak in some lines, while it was extremely high in others. We also isolated a zebrafish cytoskeletal beta-actin promoter and generated transgenic zebrafish using a beta-actin-GFP construct. In all of the four lines generated, GFP was expressed throughout the body like the beta-actin gene, demonstrating that consistent expression could also be achieved in this case. In the present study, we also examined the effects of factors which potentially affect the transgenic frequency or expression levels. The following results were obtained: (i) expression levels of GFP in the injected embryo were not strongly correlated to transgenic frequency; (ii) the effect of the NLS peptide (SV40 T antigen nuclear localization sequence), which has been suggested to facilitate the transfer of a transgene into embryonic nuclei, remained to be elusive; (iii) a plasmid vector sequence placed upstream of the construct might reduce the expression levels of the reporter gene.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
0012-1606
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
192
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
289-99
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:9441668-Actins, pubmed-meshheading:9441668-Animals, pubmed-meshheading:9441668-Animals, Genetically Modified, pubmed-meshheading:9441668-Antigens, Polyomavirus Transforming, pubmed-meshheading:9441668-Cloning, Molecular, pubmed-meshheading:9441668-Embryo, Nonmammalian, pubmed-meshheading:9441668-Gene Expression Regulation, Developmental, pubmed-meshheading:9441668-Genes, Reporter, pubmed-meshheading:9441668-Green Fluorescent Proteins, pubmed-meshheading:9441668-Luminescent Proteins, pubmed-meshheading:9441668-Muscle, Skeletal, pubmed-meshheading:9441668-Organ Specificity, pubmed-meshheading:9441668-Promoter Regions, Genetic, pubmed-meshheading:9441668-Recombinant Fusion Proteins, pubmed-meshheading:9441668-Simian virus 40, pubmed-meshheading:9441668-Transgenes, pubmed-meshheading:9441668-Zebrafish
pubmed:year
1997
pubmed:articleTitle
High-frequency generation of transgenic zebrafish which reliably express GFP in whole muscles or the whole body by using promoters of zebrafish origin.
pubmed:affiliation
PRESTO, Japan Science and Technology Corporation (JST), Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't