A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice: construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene

The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing...

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Bibliographic Details
Published in:Development genes and evolution 2000-02, Vol.210 (2), p.105-109
Main Authors: Chiu, C H, Amemiya, C T, Carr, J L, Bhargava, J, Hwang, J K, Shashikant, C S, Ruddle, F H, Wagner, G P
Format: Article
Language:English
Subjects:
Animals
Cloning
Danio rerio
DNA, Recombinant
Freshwater
Gene expression
Gene Expression Regulation, Developmental
Gene Targeting - methods
Genes, Reporter
Genetic Vectors
Homeodomain Proteins - genetics
Hoxa11b gene
Lac Operon
lacZ gene
Mice
Mice, Transgenic
Proteins
Rodents
Zebrafish - genetics
Zebrafish Proteins
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Summary:The identification of cis-sequences responsible for spatiotemporal patterns of gene expression often requires the functional analysis of large genomic regions. In this study a 100-kb zebrafish Hoxa-11b-lacZ reporter gene was constructed and expressed in transgenic mice. PAC clone 10-O19, containing a portion of the zebrafish HoxA-b cluster, was captured into the yeast-bacterial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ reporter gene was then inserted in-frame into exon 1 of the zfHoxa-11b locus by a second round of recombinogenic targeting. Expression of the zfHoxa-11b-lacZ reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only in the posterior portion of the A-P axis, in the paraxial mesoderm, neural tube, and somites. These findings demonstrate the utility of recombinogenic targeting for the modification and expression of large inserts captured from P1/PAC clones.
ISSN:0949-944X
1432-041X
DOI:10.1007/s004270050016