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.
Citation
Chiu, C.H., Amemiya, C.T., Carr, J.L., Bhargava, J., Hwang, J.K., Shashikant, C.S., Ruddle, F.H., and Wagner, G.P. (2000) 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. Development genes and evolution. 210(2):105-109.
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
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping