Journal
DEVELOPMENT GENES AND EVOLUTION
Volume 210, Issue 2, Pages 105-109Publisher
SPRINGER VERLAG
DOI: 10.1007/s004270050016
Keywords
cis-regulatory sequences; P1/PAC clones; homologous recombination; reporter gene; AbDb-like Hox genes
Funding
- NIAID NIH HHS [AI39008-01] Funding Source: Medline
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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.
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