PUBLICATION

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

Authors
Chiu, C.H., Amemiya, C.T., Carr, J.L., Bhargava, J., Hwang, J.K., Shashikant, C.S., Ruddle, F.H., and Wagner, G.P.
ID
ZDB-PUB-000308-3
Date
2000
Source
Development genes and evolution   210(2): 105-109 (Journal)
Registered Authors
Amemiya, Chris, Chiu, Chi-Hua, Ruddle, Frank H.
Keywords
cis-regulatory sequences; P1/PAC clones; homologous recombination; reporter gene; AbDb-like Hox genes
MeSH Terms
  • Animals
  • DNA, Recombinant
  • Gene Expression Regulation, Developmental
  • Gene Targeting/methods*
  • Genes, Reporter
  • Genetic Vectors
  • Homeodomain Proteins/genetics*
  • Lac Operon
  • Mice
  • Mice, Transgenic
  • Zebrafish/genetics*
  • Zebrafish Proteins*
PubMed
10664153 Full text @ Dev. Genes Evol.
Abstract
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.
Genes / Markers
Figures
Expression
Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes