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
Ch. Chiu et al., 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, DEV GENES E, 210(2), 2000, pp. 105-109
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-bact
erial shuttle vector, pPAC-ResQ, by recombinogenic targeting. A lacZ report
er 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-lac
Z reporter gene in 10.5 d.p.f. transgenic mouse embryos was observed only i
n the posterior portion of the A-P axis, in the paraxial mesoderm, neural t
ube, and somites. These findings demonstrate the utility of recombinogenic
targeting for the modification and expression of large inserts captured fro
m P1/PAC clones.