Genomic cloning of mud loach Misgurnus mizolepis (Cypriniformes, Cobitidae) beta-actin gene and usefulness of its promoter region for fish transgenesis

As a part of our effort to obtain a strong regulatory element for the construction of an autogenic mud loach transgenic vector, we isolated a genomic clone that contains an open reading frame encoding the beta-actin gene, then examined the transcriptional activity of the upstream sequences, includin...

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Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2003-05, Vol.5 (3), p.244-252
Main Authors: Noh, Jae Koo, Cho, Kyu-Nam, Han, Eun Hwa, Kim, AeRi, Lee, Jae-Seong, Kim, Dong Soo, Kim, Chul Geun
Format: Article
Language:English
Subjects:
Actins - genetics
Animals
Base Sequence
Cells, Cultured
Cloning, Molecular
Cypriniformes - genetics
Female
Freshwater
Gene Expression - genetics
Gene Transfer Techniques
Growth Hormone - genetics
Growth Hormone - physiology
Misgurnus mizolepis
Molecular Sequence Data
Promoter Regions, Genetic - genetics
Restriction Mapping
Reverse Transcriptase Polymerase Chain Reaction
Transcriptional Activation - genetics
Transfection
Transgenes - physiology
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Summary:As a part of our effort to obtain a strong regulatory element for the construction of an autogenic mud loach transgenic vector, we isolated a genomic clone that contains an open reading frame encoding the beta-actin gene, then examined the transcriptional activity of the upstream sequences, including the first intron, in transiently transfected cell lines. It showed that the upstream region has substantially strong transcriptional activity, and that both the proximal promoter and distal region of intron 1 play a crucial role in the activity. A similar result, based on fish growth, was obtained with the expression vectors containing the growth hormone gene of mud loach. These were driven by the regulatory region of the mud loach beta-actin gene with various mutations, and were directly transferred into the trunk muscle of fish using an electrostimulation-mediated method. Fish weights were monitored over the next 4 weeks. These data suggest that the proximal promoter and the first intron enhancer of the mud loach beta-actin gene are useful for autogenic mud loach transgenesis.
ISSN:1436-2228
DOI:10.1007/s10126-002-0066-1