TY - JOUR
T1 - A recombinogenic targeting method to modify large-inserts for cis-regulatory analysis in transgenic mice
T2 - construction and expression of a 100-kb, zebrafish Hoxa-11b-lacZ reporter gene
AU - Chiu, Chi Hua
AU - Amemiya, Chris T.
AU - Carr, Janet L.
AU - Bhargava, Jaya
AU - Hwang, Joseph K.
AU - Shashikant, Cooduvalli S.
AU - Ruddle, Frank H.
AU - Wagner, Günter P.
PY - 2000/1/1
Y1 - 2000/1/1
N2 - 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.
AB - 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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U2 - 10.1007/s004270050016
DO - 10.1007/s004270050016
M3 - Article
C2 - 10664153
AN - SCOPUS:0033963666
VL - 210
SP - 105
EP - 109
JO - Development Genes and Evolution
JF - Development Genes and Evolution
SN - 0949-944X
IS - 2
ER -